බເᨭㄢ㢟㸸2013㹼2015 ᖺᗘ ⬻༞୰㛵㐃㑇ఏᏊ㛵ࡍࡿ◊✲ ᇶ┙◊✲㸦㹀㸧 㰺⸨ ᘏே࣭ᮾிᏛ་Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ᡃࠎࡣ⬻᱾ሰࡢࡓࡿཎᅉ࡛࠶ࡿ㢌ෆᖿື⬦ ⊃✽㸦௨ୗ㢌ෆ⊃✽㸧㛵㐃ࡢᙉ࠸㑇ఏᏊኚ␗ࢆ ྠᐃࡋࡓ(Stroke 2012)ࠋࡇࢀࡣ≉Ⓨᛶࡢ㢌ෆ⊃✽ ࡛࠶ࡿࡶࡸࡶࡸࡢឤཷᛶ㑇ఏᏊኚ␗ࡋ࡚≉ᐃࡉ ࢀࡓ RNF213 c.14576G㸼A (rs112735431) ࡛࠶ࡿࠋ ඛ⾜◊✲࠾࠸࡚ࠊ2011 ᖺ 11 ᭶ࡽ 2014 ᖺ 4 ᭶ࡢ㛫ᮾிᏛ㝃ᒓ㝔⬻⚄⤒እ⛉࠾ࡼࡧ㛵ᮾ⬻ ⚄⤒እ⛉㝔࡚ᵝࠎ࡞⬻⾑⟶ᝈࡢᝈ⪅ 649 㸦ࡶࡸࡶࡸ 91 ࠊ∦ഃࡶࡸࡶࡸ 19 ࠊ㢌ෆ ⊃✽ 185 ࠊ㢁ື⬦⊃✽ 74 ࠊ⬻ື⬦⒗ 110 ࠊ 㧗⾑ᅽᛶ⬻ෆฟ⾑ 25 ࠊnormal control 145 㸧 ࠾࠸࡚ RNF213 c.14576G㸼A ࡢ᭷↓ࢆ᳨ドࡋࡓࠋ ኚ␗ࢆᣢࡘྜࡣࡶࡸࡶࡸ࡛ࡣ 84.6%㸦77/91㸧ࠊ ∦ഃ࡛ࡣ 63.1%㸦12/19㸧࡛࠶ࡗࡓࠋࡉࡽࡣࢸ ࣟ ࣮ ࣒ ◳ ᛶ ࡢ 㢌 ෆ ⊃ ✽ ࠾ ࠸ ࡚ ࡣ 24.3% 㸦45/185㸧࡛࠶ࡗࡓࠋࡑࡢࡢ⩌࡛ࡣ㢁ື⬦⊃✽ 1 ࠊ⬻ື⬦⒗ 1 ࠊcontrol 2 ㄆࡵࡿࡢࡳ࡛࠶ ࡗࡓࠋc.14576G㸼A ࡣࡶࡸࡶࡸࡤࡾ࡛࡞ࡃࠊື ⬦◳ᛶࡢ㢌ෆ⊃✽ᑐࡋ࡚ࡶ᭷ព࡞㛵㐃ࡀ♧ࡉ ࢀࡓ(Stroke 2013)ࠋ ࡇࡢኚ␗ࡀ୍⯡ࡢື⬦◳ᛶࡢ㢌ෆ⊃✽ࢆࡁࡓ ࡍ㑇ఏⓗࣜࢫࢡࣇࢡࢱ࣮࡛࠶ࡿ⪃࠼ከゅⓗ࡞᳨ ウࢆ⾜࠺ࡇࢆᮏ◊✲ࡢ┠ⓗࡋࡓࠋ 㸺2013 ᖺᗘࡢᨭࡢෆᐜ㸼 ඛ⾜◊✲࠾࠸࡚ RNF213 ࡀ⬻༞୰㛵㐃㑇ఏᏊࡋ ࡚㔜せ࡛࠶ࡿࡇࢆ᫂ࡽࡋࡓࠋ୍᪉࡛ࠊ㢌ෆ ⊃✽ࢆ᭷ࡍࡿᝈ⪅ࡢ୰ࡣኚ␗ c.14576G>A ࢆᣢࡓ ࡞࠸⪅ࡶᏑᅾࡍࡿࠋࡇࢀࡽࡢᝈ⪅ࡢ୰ࡣ c.14576G>A ௨እࡢ RNF213 ࡢኚ␗ࢆᣢࡘ⪅ࡀᏑᅾ ࡍࡿྍ⬟ᛶࡀ࠶ࡿࠋࡑࡇ࡛ࠊࡇࢀࡲ࡛ RNF213 ࡢ㑇 ఏᏊኚ␗ c.14576G>A ࡢࡳ╔┠ࡋゎᯒࢆ⾜ࡗ࡚ࡁ ࡓࡀࠊᝈⓎ㛵㐃ࡍࡿ RNF213 ୖࡢࡑࡢࡢኚ ␗ࡢྠᐃࢆ┠ᣦࡋࡓࠋ㢌ෆ⊃✽ᝈ⪅ 271 ேᖖ ⪅ 141 ே࠾࠸࡚ RNF213 యࡢ resequence ࢆ⾜ ࠸ࠊc.14576G>A ௨እࡢᝈ㛵㐃ࡍࡿኚ␗ྠᐃࢆ ┠ᣦࡋࡓࠋRNF213 ࡣ exon ᩘࡀ 68 ࡁ࡞㑇ఏᏊ ࡛ ࠶ ࡾ ࠊ ḟ ୡ ௦ ࢩ ࣮ ࢡ ࢚ ࣥ ࢧ ࣮ ࢆ ⏝ ࠸ ࡓ target resequence ࢆ⾜ࡋ࡚㡬࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ゎᯒࡢ⤖ᯝ RNF213 ୖ 185 ࡢኚ␗㸦ከᆺ㸧ࡀྠ ᐃࡉࢀࡓࠋࡑࡢෆヂࡋ࡚ࡣࠊ82 ࡀ Noncoding 㡿ᇦ ࡛ࠊ103 ࡀ Coding 㡿ᇦᏑᅾࡋࡓࠋCoding 㡿ᇦࡢ ኚ␗㸦ከᆺ㸧ࡢෆヂࡣ Synonymous 42ࠊMissense ࡀ 61ࠊNonsense 0ࠊFrameshift 0 ࡛࠶ࡗࡓࠋ Coding 㡿ᇦࡢኚ␗㸦ከᆺ㸧ࡢ࠺ࡕࠊᝈⓎ㛵 ࢃࡿࡶࡢࡣ Missense ࡢࡶࡢ⪃࠼ࠊ61 ࡢ Missense ࡢኚ␗㸦ከᆺ㸧࠾࠸࡚ Case-control study ࢆ⾜࠸ࠊ case ᭷ព࡞㛵㐃ࡀ࠶ࡿኚ␗㸦ከᆺ㸧ࡢྠᐃࢆ┠ᣦ ࡋࡓࠋ⤖ᯝࡋ࡚ࡣࡇࢀࡲ࡛╔┠ࡋ࡚ࡁࡓ c.14576G 㸼A (rs112735431)ࡀ᭱ࡶᝈᙉ࠸㛵㐃ࢆᣢࡘኚ ␗࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ࡲࡓᝈ⩌࡛ࡢࡳㄆࡵࡽࢀࡿ rare variant ࡀ 8 ✀ 㢮ྠᐃࡉࢀࠊc.14576G㸼A (rs112735431)௨እ࡛ᝈ 㛵㐃ࡍࡿ㑇ఏᏊኚ␗ࡢೃ⿵࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ௨ୖࡢ⤖ᯝࡼࡾࠊࡇࢀࡲ࡛╔┠ࡋ࡚ࡁࡓ㑇ఏᏊኚ ␗ RNF213 c.14576G>A ࡢ㔜せᛶࡀ☜ㄆࡉࢀࡿ ⤖ᯝ࡞ࡗࡓࠋࡲࡓ RNF213 c.14576G>A ࢆࡶࡓ࡞ ࠸Ⓨ⪅ࡇࢀࡲ࡛ሗ࿌ࡢ࡞࠸᪂つࡢኚ␗ࢆᩘ✀㢮 ㄆࡵࡽࢀࠊᝈ㛵ࢃࡿ㑇ఏᏊኚ␗ࡢೃ⿵⪃࠼ࡽ ࢀࡿࠋࡇࢀࡽࡢೃ⿵࡞ࡿኚ␗㛵ࡋ࡚ࡣูࢥ࣮࣍ ࢺ࡛ࡢ validation ࡸኚ␗ࡢ⏕≀Ꮫⓗព⩏ࡢ᳨ドࡀᚲ せ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋ 㸺2014, 2015 ᖺᗘࡢᨭࡢෆᐜ㸼 ௨ୖࡢ⤖ᯝࡽᡃࠎࡣ RNF213 c.14576G>A ࡀ⬻ ༞୰ࡢࡓࡿཎᅉ࡛࠶ࡿ㢌ෆ⊃✽ࡁ࡞㛵㐃ࢆ ᣢ ࡘ ࡇ ࢆ ᫂ ࡽ ࡋ ࡓ ࠋ RNF213 ኚ ␗ 㸦c.14576G>A㸧ࡣ᪥ᮏேࡢ୍⯡㞟ᅋࡢ 2%ẚ㍑ⓗ 㧗㢖ᗘㄆࡵࡽࢀࡿࡀ᫂ࡽ࡞ࡗ࡚࠾ࡾࠊ୍᪉࡛ 㢌ෆ⊃✽ࡢ᭷⋡ࡣ⣙ 1000 ே 1 ே⛬ᗘ㸦యࡢ 0.1%⛬ᗘ㸧࠶ࡾࠊRNF213 c.14576G㸼A ࡢಖᅉ⪅㸦 యࡢ 2%⛬ᗘ㸧ࡀࡍ࡚Ⓨࡍࡿࢃࡅ࡛ࡣ࡞࠸ࠋࡘࡲ ࡾ RNF213 c.14576G㸼A ࢆᣢࡗ࡚࠸࡚ࡶࠊ㢌ෆ⊃ ✽ࡢⓎࡢ᭷↓ࠊ㔜ᗘ㛵ࢃࡿࡉࡽูࡢ㑇ఏᏊኚ ␗ ࡀᏑᅾ ࡍࡿྍ ⬟ᛶ ࡀ㧗࠸ ⪃࠼ ࡽࢀࡿ ࠋ 2014, 2015 ᖺ࠾࠸࡚ࡣࡇࡢࡼ࠺࡞ᝈⓎࡢỴᐃᅉᏊ ࡞ࡿ᪂ࡓ࡞㑇ఏⓗኚ␗㸦㑇ఏᏊከᆺ㸧ࢆ᫂ࡽ ࡍࡿࡇࢆ┠ⓗࡋࡓࠋ 2014 ᖺࡣᐙ⣔ゎᯒ⾜ࡗࡓࠋࡍ࡞ࢃࡕྠ୍ᐙ⣔ෆ࡛ RNF213 c.14576G㸼A ࢆ᭷ࡋ࡞ࡀࡽⓎ⪅ࠊ㠀Ⓨ ⪅ࢆㄆࡵࡿᐙ⣔ࢆ 15 ⣔ 30 ᳨యࡢ whole exome ゎᯒ ࢆ⾜࠸ࠊ㑇ఏᏛⓗᡭἲࢆ⏝࠸࡚ࢤࣀ࣒㡿ᇦࡽ ᝈⓎࡢỴᐃᅉᏊ࡞ࡿ㑇ఏᏊኚ␗ࡢྠᐃࢆヨࡳ࡚ ࠸ࡿࠋ2015 ᖺᗘࡣ RNF213 c.14576G㸼A ࢆ᭷ࡍ ࡿⓎ⪅⩌㠀Ⓨ⪅⩌ࡢ 2 ⩌㛫࡛ whole exome ゎ ᯒࢆ⾜࠺᪉㔪ࡋࡓࠋⓎ⪅ࢆ 100 ே whole exome ゎᯒࢆ⾜࠸ࠊࡲࡓ 76 ྡࡢ㠀Ⓨ⪅㸦ᮾிᏛࢤࣀ࣒ ་Ꮫࢭࣥࢱ࣮ࡢࢹ࣮ࢱ࣮࣋ࢫࡢࢹ࣮ࢱࢆ⏝㸧㛵 㐃ゎᯒࢆ⾜ࡗ࡚࠸ࡿࠋ࠸ࡎࢀࡢゎᯒࡶ⌧ᅾ㐍⾜୰࡛ ࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 RNF213 c.14576G㸼A ࡣᵝࠎ࡞⾲⌧ᆺࡢ㢌ෆ⊃ ✽᭷ព࡞㛵㐃ࡀ♧ࡉࢀࠊࡲࡓࡇࡢኚ␗ࡣ᪥ᮏேࡢ 2%⛬ᗘࡀࡶࡘࡇࡀ☜ㄆ࡛ࡁࡓࠋࡇࡢኚ␗ࡣ㧗⾑ᅽ ࡢ㛵㐃ࡍࡿࡇࡶሗ࿌ࡉࢀ࡚࠾ࡾࠊ㢌ෆ⊃✽ࡢ ࣜࢫࢡࣞࣝ┦ࡲࡗ࡚⬻༞୰㛵㐃㑇ఏᏊኚ␗㸦ࣂ ࣐࣮࣮࢜࢝ೃ⿵㸧ࡽ࠼ࡿࡇࡶ࡛ࡁࡿࠋࡇࡢ ኚ␗ࢆࡶࡘࣜࢫࢡ⩌ᑐࡋ࡚ࡣࠊMRI ࡞ࡢ⬻⢭ᰝ ࡸ᪩ᮇࡽࡢ㝆ᅽ⸆⒪ධ࡞ࢆ✚ᴟⓗ࠾ࡇ࡞ ࠺ࡇࡼࡾࠊ⬻༞୰ࡢண㜵་⒪ࡍ࡞ࢃࡕඛไ་⒪ ࡘ࡞ࡀࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 353 - බເᨭㄢ㢟㸸2011 ᖺᗘ ඹ⏕㑇ఏᏊ⩌᭱ᑠࢭࢵࢺࡢྠᐃࢆ┠ᣦࡍ࣑ࣖࢥࢢࢧ᰿⢏⳦ඹ⏕ࣛࣥࢻࡢ᭱ᑠ ᇶ┙◊✲㸦㹁㸧 బ ᙪ࣭ዉⰋዪᏊᏛ ◊✲㝔 ⮬↛⛉Ꮫ⣔࣭⏕≀⛉Ꮫ㡿ᇦ 㸺⫼ᬒ࣭┠ⓗ㸼 ᰿⢏⳦࣐࣓⛉᳜≀ࡀႠࡴ❅⣲ᅛᐃඹ⏕ࡣࠊ┦ඹ ⏕ࡢ௦⾲ⓗ࡞࡛࠶ࡾࠊᩘᑡ࡞࠸⣽⬊ෆඹ⏕ࡢᐇ ࡛ࡶ࠶ࡿࠋ␗✀⏕≀㛫ࡢ┦ㄆ㆑ᐶᐜ࣭㝖ࡢᶵ ᵓࠊࡉࡽࡑࡢ㐍ࡣᇶ♏⛉Ꮫୖࡢ㔜せ࡞ㄢ㢟࡛࠶ ࡿࡶࠊࢲࢬ࡞ࡢ㔜せస≀⏕⏘ᛶᨵⰋࡢᇶ ┙࡞ࡿㄢ㢟ゝ࠼ࡿࠋඹ⏕ᚲせ࡞㑇ఏᏊࡢ᭱ᑠ ࢭࢵࢺࢆྠᐃࡍࡿࡇࡣࠊᨵⰋࡢ㐨➽ࢆ♧ࡍୖ࡛㔜 せ࡛࠶ࡿࠋ ᰿⢏⳦ഃࡢඹ⏕㑇ఏᏊࡋ࡚ࡣࠊ᰿⢏ࡢᙧᡂᶵ⬟ ࡍࡿ㑇ఏᏊ⩌㸦nod㸧ࠊ❅⣲ᅛᐃᛂࢆ⾜࠺ࢽࢺࣟࢤ ࢼ࣮ࢮࡑࡢᡂ⇍࡞㛵ࡍࡿ❅⣲ᅛᐃ㑇ఏᏊ⩌ 㸦nif㸧ࢽࢺࣟࢤࢼ࣮ࢮ௨እ࡛᰿⢏ෆ࡛ࡢ❅⣲ᅛᐃ 㛵ࡍࡿ㑇ఏᏊ⩌㸦fix㸧࡞ຍ࠼࡚ࠊࡉࡽᩘ ከࡃࡢ㑇ఏᏊ⩌ࡀᚲせ࡛࠶ࡿࠋ᪤▱ࡢ᰿⢏⳦ࡣࠊ㏆ ⦕ࡢ㠀᰿⢏⳦ඹ㏻ࡍࡿࢤࣀ࣒ࡢࢭࢵࢺຍ࠼࡚ࠊ ࢤࣀ࣒ୖࡢ୍㒊እ᮶ᛶࡢࣛࣥࢻࡋ࡚ࠊ࠶ ࡿ࠸ࡣࣉࣛࢫ࣑ࢻࡋ࡚ୖグࡢ nod, nif fix 㑇 ఏᏊ⩌ࢆಖᣢࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋࡋࡋࠊඹ ⏕ᚲせ࡞㑇ఏᏊࡢ᭱ᑠࢭࢵࢺࡢᐇయࡣᮍࡔᮍゎ᫂ ࡢㄢ㢟࡛࠶ࡿࠋᮏ◊✲࡛ࡣࠊ࣐࣓⛉ࡢࣔࢹ᳜ࣝ≀࡛ ࠶ࡿ࣑ࣖࢥࢢࢧࡢ᰿⢏⳦ Mesorhizobium loti ࡢඹ ⏕ࣛࣥࢻࡘ࠸࡚ࠊ␗࡞ࡿ⳦ᰴ࣭㔝⏕⣔⤫ࡢ㓄 ิࢹ࣮ࢱᇶ࡙࠸࡚Ḟኻࢆᑟධࠊ᭱ᑠඹ⏕ࣛࣥ ࢻࢆᐇయࡋࠊඹ⏕㑇ఏᏊ⩌ࡢ᭱ᑠࢭࢵࢺࡢゎ᫂ ㏕ࡿࡇࢆ┠ⓗࡍࡿࠋࡲࡓࠊࡇࡢ┠ⓗ㐩ᡂ㝶 ࡋ࡚ࠊ᰿⢏⳦ࡑࡢ㏆⦕✀࡛⏝ྍ⬟࡞㑇ఏᏊᕤᏛ ࢶ࣮ࣝ࢟ࢵࢺ㸦㑇ఏᏊⓎ⌧◚ቯ⏝㸧ࡢᩚഛࡶ⾜࠺ࠋ 㸺ᨭࡢෆᐜ㸼 ࣑ࣖࢥࢢࢧ᰿⢏⳦࣭㔝⏕⣔⤫㸲ᰴࡢ Roche 454 ࡼ ࡿࠊࢻࣛࣇࢺ࣭ࢩ࣮ࢣࣥࢫゎᯒ㸸㸲ᰴࡢࢤࣀ࣒ࢧ ࢬࡣࠊ6.7 Mbpࠥ7.3 Mbp ࡛ࠊ17.0 x ࡽ 44.7 x ࢝ ࣂ࣮ࡼࡾࠊ2 ࡽ 8 ࡢ scaffold ࢵࢭࣥࣈࣝ㸦ᮾ ிᏛᏛ㝔᪂㡿ᇦᡂ⛉Ꮫ◊✲⛉㸦ᓥᚿᮁඛ ⏕ࠊ᭹㒊ṇᖹඛ⏕㸧ࡼࡿ㸧ࠋ ୖグࡣูࡢ࣑ࣖࢥࢢࢧ᰿⢏⳦࣭㔝⏕⣔⤫㸶ᰴࡢࠊ HiSeq ࣓ࢺ࣌ࢹ࣮ࢱ㸦3kbp 4kbp ࡛ 101 ࢧ ࢡࣝ㸧ࠊࡇࢀࡽࡢ࠺ࡕ 5 ᰴࡘ࠸࡚ MiSeq ࢚࣌ࣥ ࢻࢹ࣮ࢱ㸦400 bp ࡛ࠊ150 ࢧࢡࣝ㸧ྲྀᚓ㸦ᅜ❧㑇 ఏᏛ◊✲ᡤ㸦㇏⏣ᩔඛ⏕㸧ࡼࡿ㸧 ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣑ࣖࢥࢢࢧ᰿⢏⳦ࡢඹ⏕ࣛࣥࢻ࣭ࢥ㑇ఏᏊࡢ ☜ᐃ㸸࣑ࣖࢥࢢࢧ᰿⢏⳦㔝⏕⣔⤫㸲ᰴࡢ 454 ゎᯒࢹ ࣮ࢱࡽࠊ࠸ࡎࢀࡢ⳦ᰴ࡛ࡶ Phe-tRNA 㑇ఏᏊࢆศ ᩿⏕ࡉࡏࡿᙧᘧ࡛ඹ⏕ࣛࣥࢻࡀỈᖹఏᦙࡋ࡚ ࠸ࡿࡇࡀ♧ࡉࢀࡓࠋࣛࣥࢻࡢࢧࢬࡣ 415 ࡽ 615kbp ࡛ࠊ400ࠥ600 ⛬ᗘࡢ ORF ࢆྵࢇ࡛࠸ࡓࠋ HiSeq MiSeq ゎᯒࢆ⾜ࡗࡓࡢ⳦ᰴࡶྵࡵ࡚ࠊ࠸ ࡎࢀࡢඹ⏕ࣛࣥࢻࡶඹ㏻ࡋ࡚ྵࡲࢀࡿࢥ㑇 ఏᏊᩘࡣ 165 ࡛࠶ࡗࡓࠋࡇࢀࡽࡢࢥ㑇ఏᏊࡣ᰿⢏ ࡢᙧᡂᶵ⬟ࡍࡿ㑇ఏᏊ⩌㸦nod㸧ࠊ❅⣲ᅛᐃᛂࢆ ⾜࠺ࢽࢺࣟࢤࢼ࣮ࢮࡑࡢᡂ⇍࡞㛵ࡍࡿ❅⣲ ᅛᐃ㑇ఏᏊ⩌㸦nif㸧ࢽࢺࣟࢤࢼ࣮ࢮ௨እ࡛᰿⢏ෆ ࡛ࡢ❅⣲ᅛᐃ㛵ࡍࡿ㑇ఏᏊ⩌㸦fix㸧࠾ࡼࡧඹ⏕ ࣛࣥࢻࡢỈᖹఏᦙ㛵ࢃࡿ trb 㑇ఏᏊ࡞ࢆྵ ࡴᴫࡡ㸱ࡘࡢࢡࣛࢫࢱ࣮ศࢀࠊ㸱ࡘࡢࢡࣛࢫࢱ ࣮㛫ࡢ㓄ิࡣⴭࡋࡃ㧗࠸ከᵝᛶࡀㄆࡵࡽࢀࡓࠋ ࣑ࣖࢥࢢࢧ᰿⢏⳦ࡢᰁⰍయࡢከᵝᛶ㸸࣑ࣖࢥࢢࢧ ᰿⢏⳦ࢤࣀ࣒ࡢඹ⏕ࣛࣥࢻ௨እࡢ㡿ᇦࡣࠊయ ࡋ࡚┦ಖᏑࡣㄆࡵࡽࢀࡿࡶࡢࡢࠊ࠼ࡤ 16S-23S rRNA 㓄ิࡢከᵝᛶྠᵝࡢከᵝᛶࡀㄆࡵ ࡽࢀࡓࠋඹ⏕ࣛࣥࢻࡢᤄධࡉࢀ࡚࠸ࡿ PhetRNA ௨እࡶ」ᩘࡢ rRNA 㑇ఏᏊࡢᤄධᅉᏊࡢ Ꮡᅾࡀㄆࡵࡽࢀࡓࠋ⳦ᰴ㛫ࡢẚ㍑ࡼࡾࠊᰁⰍయ ഃࡢࢥ㑇ఏᏊࡢᴫせࡀุ᫂ࡋࡓࠋ ࣑ࣖࢥࢢࢧ᰿⢏⳦㔝⏕ᰴࡢ⪏ሷᛶ㑇ఏᏊ㸸࣑ࣖࢥࢢ ࢧ᰿⢏⳦㔝⏕⣔⤫ࡣ㸱㸣⛬ᗘࡢ NaCl ࡢᏑᅾୗ࡛ ࡶάⓎቑṪࡍࡿࡶࡢࡀ」ᩘ▱ࡽࢀ࡚࠸ࡿࠋࡇࢀࡽ ࡢሷ⪏ᛶᰴ≉᭷࡞㑇ఏᏊࢭࢵࢺࡀぢࡘࡗࡓࠋሷ ⪏ᛶᰴ࡛ࡢ㑇ఏᏊ◚ቯሷឤཷᛶᰴࡢ㑇ఏᏊᑟධ ࢆ⏝࠸ࡓᶵ⬟ࡢᐇドゎᯒࢆᐇࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣐࣓⛉ࡢࣔࢹ᳜ࣝ≀࣑ࣖࢥࢢࢧࡢ᰿⢏⳦ࡣࠊᡤㅝ Mesorhizobioum ᒓࢆ㉺࠼ Aminobacter ᒓ➼ࢆྵࡳࠊ ඹ ㏻ ࡢ ♽ ඛ ࢆ ᣢ ࡘ ඹ ⏕ ࣛ ࣥ ࢻ ࡀ Phylobacteriaceae ᒓࡍࡿ⣽⳦ࡢᰁⰍయᤄධ ࡍࡿࡇࡼࡾᡂ❧ࡋ࡚࠸ࡿ᥎ᐃࡉࢀࡓࠋࡇࢀࡽ ࡢඹ⏕ࣛࣥࢻࡣࠊඹ⏕ࡣ┤᥋㛵ಀࡋ࡞࠸ࣅ ࢜ࢳࣥࡸࢳ࣑ࣥ࡞ࡢࣅࢱ࣑ࣥྜᡂ⣔㑇ఏᏊ⩌ࢆ ྵࡳࠊᰁⰍయࡢᶵ⬟ࢆ⿵࠺ᶵ⬟ࢆᣢࡕࠊ400kbp ௨ ୖࡢࢧࢬࢆᣢࡘࠗᕫⓗ㑇ఏᏊࢡࣛࢫࢱ࣮࠘ࡋ ࡚ࠊ୍ᐃࡢ⣽⳦ࡢᰁⰍయ┦స⏝ࡋࠊ᪤⋓ᚓ ῭ࡳࡢ᰿⢏ᙧᡂࡸ❅⣲ᅛᐃ㑇ఏᏊ⩌ຍ࠼࡚ࠊᰁ Ⰽయୖ⨨ࡍࡿ㑇ఏᏊ⩌ࢆྲྀᤞ㑅ᢥࡋࡘࡘ㐍ࡋ ࡚࠸ࡿࡶࡢ⪃࠼ࡽࢀࡓࠋ☜ㄆࡉࢀࡓඹ⏕ࣛࣥ ࢻࡣ࠸ࡎࢀࡶࠊࣛࣥࢻෆ⨨ࡍࡿ P4 ࣇ࣮ࢪ ᆺࣥࢸࢢ࣮ࣛࢮࡼࡾࠊPhe-tRNA 㑇ఏᏊࢆᶆⓗ ࡋࡓุ᩿ࡉࢀࠊ࣑ࣖࢥࢢࢧ᰿⢏⳦ࡢᡂ❧㐍 㛵ࡋ࡚ࠊ⤫ྜⓗ࡞⌮ゎࡀᚓࡽࢀࠊඹ⏕㑇ఏᏊࡢ᭱ ᑠࢭࢵࢺࡢᐇドⓗྠᐃࡢぢ㏻ࡋࡀࡘ࠸ࡓࠋ ࢤࣀ࣒㒊ศࡢゎᯒࡽࠊ᰿⢏⳦ࡢሷࢫࢺࣞࢫ⪏ᛶࠊ ⇱⪏ᛶ࡞ࡢཎᅉೃ⿵ࡀᢳฟ࡛ࡁࠊ⫱✀ྥࡅ࡚ ࡢᒎᮃࡀᣅࡅࡓ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Kasai-Maita H et al. Microbes and Environments 28(2): 275-278 (2013) - 354 - බເᨭㄢ㢟㸸2010ࠊ2015 ᖺᗘ ࢩࣕࢪࢡࣔ⸴㢮ࡢẚ㍑ࢤࣀ࣒㑇ఏᏊᶵ⬟ゎᯒࡽ᥈ࡿ᳜≀ࡢከ⣽⬊యไ㐍 ᇶ┙◊✲㸦㹁㸧 ᆏᒣ ⱥಇ࣭⚄ᡞᏛᏛ㝔⌮Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 㝣ୖ᳜≀ࡣ⣙ 4.8 ൨ᖺ๓Ỉ୰⏕άࢆࡍࡿ⸴㢮 ࡽ㐍ࡋࡓࠋࡇࢀࡲ࡛ࡢᙧែᏛⓗ◊✲ศᏊ⣔⤫Ꮫ ⓗ◊✲ࡢ⤖ᯝࡽࠊࢩࣕࢪࢡࣔ⸴㢮 (ࢩࣕࢪࢡࣔࠊ ࢥࣞ࢜ࢣ࣮ࢸࠊࣄ࣓࣑࢝ࢷ࢟ࣔࠊࢡࣞࣈࢯ࣑ࣝࢹ ࣒࢘➼) ࡀ㝣ୖ᳜≀᭱ࡶ㏆⦕ࡔ⪃࠼ࡽࢀ࡚࠸ࡿࠋ 㝣ୖ᳜≀ࡣࡍ࡚⬊Ꮚయࡼࡤࢀࡿ㸰ಸయከ⣽⬊ య㓄അయࡼࡤࢀࡿ㸯ಸయከ⣽⬊యࡽ࡞ࡿୡ௦ ௦ࢆ⾜ࡗ࡚࠾ࡾࠊ㝣ୖ᳜≀ࡢ㸯ಸయ㸰ಸయࡣ ࡕࡽࡶከ⣽⬊࡛࠶ࡿࠋ࠸ࡗࡱ࠺ࠊࢩࣕࢪࢡࣔ⸴㢮࡛ ࡣ㸯ಸయከ⣽⬊యࡣ࠶ࡿࡀ㸰ಸయከ⣽⬊యࡣ࡞࠸ࠋ ࡛ࡣࠊ㝣ୖ᳜≀࠾ࡼࡧྛࢩࣕࢪࢡࣔ⸴㢮ࡢከ⣽⬊య ไࡢⓎ⏕ࢩࢫࢸ࣒ࡢࡼ࠺࡞㐪࠸ࡀ࠶ࡿࡢ࡛࠶ࢁ ࠺ࠋ⏕≀㐍࠾࠸࡚ࠊ᪂つ⋓ᚓࡋࡓ㑇ఏᏊࡢ ከᵝࡀࠊࡑࡢᚋࡢᙧែ㐍ࡁࡃᐤࡋࡓ⪃ ࠼ࡽࢀ࡚࠸ࡿࠋࡇࢀࡲ࡛ࠊせ࡞㝣ୖ᳜≀ࡢᴫせ ࢤࣀ࣒ゎㄞࡀࡋࠊࡈࡃ᭱㏆ࠊࢡࣞࣈࢯ࣑ࣝࢹ ࣒࢘ࡢᴫせࢤࣀ࣒ࡀゎㄞࡉࢀࡓࡇࡽ᳜≀ࡢከᵝ ᛶࡣࢤࣀ࣒࡛ࣞ࣋ࣝẚ㍑࡛ࡁࡿࡼ࠺࡞ࡗ࡚ࡁࡓࠋ ࢩࣕࢪࢡࣔ⸴㢮࡛ࡣࠊඛ⾜◊✲ࡼࡾࠊ࠸ࡃࡘࡢ 㝣ୖ᳜≀≉␗ⓗ࡞Ⓨ⏕㑇ఏᏊࡀࢩࣕࢪࢡࣔ⸴㢮ࡶ Ꮡᅾࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ ᮏ◊✲࡛ࡣ᳜≀ከ⣽⬊యไࡢ㉳※㐍㐣⛬ࢆゎ ᫂ࡍࡿࡓࡵࠊ㝣ୖ᳜≀ࢩࣕࢪࢡࣔ⸴㢮ࡀᣢࡘ㸯ಸ యከ⣽⬊యไ㸰ಸయከ⣽⬊యไࢆ㑇ఏᏊሗࡽ ẚ㍑ࡍࡿࠋࢩࣕࢪࢡࣔ⸴㢮࠾࠸࡚⏦ㄳ⪅ࡽࡀ☜❧ ࡋࡓᇵ㣴⣔ࢆᇶ♏ࡋࠊࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒/ࢤࣀ ࣒ゎᯒࡼࡾⓎ⏕㛵ࢃࡿ㑇ఏᏊࢆ⥙⨶ⓗ㞟ࡍ ࡿࠋࡢ㑇ఏᏊࡀ㝣ୖ᳜≀ྛࢩࣕࢪࢡࣔ⸴㢮ࡢ ඹ㏻♽ඛᏑᅾࡍࡿࠊᏑᅾࡍࡿሙྜࡢࡼ࠺࡞ ᙺࢆᯝࡓࡋ࡚࠸ࡿࢆゎ᫂ࡋࠊከ⣽⬊యไ⋓ᚓ๓ ᚋ࡛ࡢ㑇ఏᏊ㐍ࢆ≉ᐃࡍࡿࠋ ࣛࣥࢫࢡࣜࣉࢺ࣮࣒ሗࢆྵࡵࠊ㝣ୖ᳜≀ࡢࢤࣀ࣒ ࡢẚ㍑ゎᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊࢩࣕࢪࢡࣔ⸴㢮 㝣ୖ᳜≀ࡢඹ㏻♽ඛࡢẁ㝵࡛ 6,300 ๓ᚋࡢ㑇ఏᏊ ࢆඹ᭷ࡋ࡚࠸ࡓ᥎ᐃࡉࢀࡓࠋࡲࡓࠊ᳜≀≉␗ⓗࡔ ⪃࠼ࡽࢀ࡚࠸ࡓከࡃࡢⓎ⏕㑇ఏᏊࡢ࣍ࣔࣟࢢࢆࢩ ࣕࢪࢡࣔ⸴㢮ࡽぢ࠸ࡔࡋࡓࠋ࠸ࡗࡱ࠺ࠊࢩࣕࢪࢡ ࣔ⸴㢮ࡢྛ⣔⤫≉␗ⓗ࡞㑇ఏᏊࡢᩘࡀከ࠸ࡇࡸࠊ ࢩࣟࢾࢼࢬࢼࢾ࢝ࢱࣄࣂᏑᅾࡋࠊࣄ࣓ࢶࣜ ࢞ࢿࢦࢣᏑᅾࡋ࡞࠸⪃࠼ࡽࢀ࡚࠸ࡓ࠸ࡃࡘࡢ Ⓨ⏕㑇ఏᏊࡀࢩࣕࢪࢡࣔ⸴㢮Ꮡᅾࡍࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡲࡓࠊࢩࣕࢪࢡࣔ⸴㢮㝣ୖ᳜≀ࡀඹ ᭷ࡍࡿⓎ⏕㑇ఏᏊࡘ࠸࡚㑇ఏᏊ⣔⤫ᶞࢆᵓ⠏ࡋࠊ ┦ྠ㑇ఏᏊ㛫ࡢ⣔⤫㛵ಀ㑇ఏᏊ㔜」ࣃࢱ࣮ࣥࢆゎ ᯒࡋࡓࠋࡑࡢ⤖ᯝࠊࢩࣕࢪࢡࣔ⸴㢮ࡢ㑇ఏᏊࡣࠊྛ 㑇ఏᏊࣇ࣑࣮ࣜࡸࡑࡢࢧࣈࣇ࣑࣮ࣜ࠾࠸࡚⣔ ⤫ⓗᇶ㒊⨨ࡍࡿࡇࡸࠊ㝣ୖ᳜≀ࡢẁ㝵࡛ከ ᵝࡀ㐍ࢇ࡛࠸ࡿ㑇ఏᏊࣇ࣑࣮ࣜࡢከࡃࡣࠊࢩࣕ ࢪࢡࣔ⸴㢮ࡢẁ㝵࡛ࡣ㛗ᛶࡀపࡃከᵝࡀ㐍ࢇ࡛ ࠸࡞࠸ࡇࡀ☜ㄆࡉࢀࡓࠋ ࠸ࡃࡘࡢ㑇ఏᏊࡘ࠸࡚ࠊࢩࣕࢪࢡࣔ࠾ࡅࡿ Ⓨ⌧ᵝᘧࢆ RT-PCR ἲࠊin situ ࣁࣈࣜࢲࢮ࣮ࢩࣙ ࣥἲࡼࡗ࡚ゎᯒࡋࠊⓎ⏕ẁ㝵ࡸჾᐁ/⣽⬊ẖ≉␗ ⓗⓎ⌧ࡍࡿ㑇ఏᏊࢆ≉ᐃࡋࡓࠋࡲࡓࠊࢩࣕࢪࢡࣔ ࡢ㑇ఏᏊࢆࢩࣟࢾࢼࢬࢼᑟධࡋᶵ⬟ゎᯒࢆ⾜࠸ࠊ 㑇ఏᏊࡢᵓ㐀ࡸᶵ⬟ࠊ࠾ࡼࡧࢩࣕࢪࢡࣔ⸴㢮㝣ୖ ᳜≀ࡀᣢࡘከ⣽⬊యไࡢ㐍㐣⛬ࢆ⪃ᐹࡋࡓࠋࡉࡽ ࠊࢩࣕࢪࢡࣔࡢࢤࣀ࣒ሗᇶ࡙ࡁࠊ⏕ά⎔ࢆ㏻ ࡋ࡚㧗࠸Ⓨ⌧ࢆ♧ࡍ⪃࠼ࡽࢀࡿ㑇ఏᏊࡢࣉ࣮ࣟࣔ ࢱ࣮ࡀྵࡲࢀࡿࢤࣀ࣒㡿ᇦࢆ⏝࠸ࠊእ᮶㑇ఏᏊࢆⓎ ⌧ࡉࡏࡿࢥࣥࢫࢺࣛࢡࢺࢆస〇ࡋࠊ㑇ఏᏊᑟධࡢ᮲ ௳᳨ウࢆ㐍ࡵࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࣭ࢩࣕࢪࢡࣔ㸦Chara braunii, S276 ᰴ㸧࠾࠸࡚ࠊᵝࠎ ࡞ࢫࢸ࣮ࢪ࠶ࡿ⣽⬊ࡽ 8 ✀ࡢ RNA ࢆㄪᩚࡋࠊḟ ୡ௦ࢩ࣮ࢣࣥࢧ࣮㸦Illumina㸧ࢆ⏝࠸ࡓ RNA-seq ゎᯒ ࢆᮏᨭࡢࡶᐇࡋࡓ㸦2010 ᖺᗘ㸧 ࠋ ࣭ࢩࣕࢪࢡࣔࡢჾᐁ/⣽⬊ẖࠊⓎ⏕ࢫࢸ࣮ࢪẖࡢ RNA 㸦ィ 12 ✀ࠊྛ 3 ᳨య㸧ࢆㄪᩚࡋࠊḟୡ௦ࢩ࣮ࢣࣥࢧ ࣮㸦Illumina㸧ࢆ⏝࠸ࡓ RNA-seq ゎᯒࢆᮏᨭࡢࡶ 㐍ࡵ࡚࠸ࡿ㸦2015 ᖺᗘ㸧 ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ௨ୖࡢ◊✲ࢆ㏻ࡋ࡚ࠊࢩࣕࢪࢡࣔ⸴㢮ࡢࢤࣀ࣒ࡢ ≉ᚩࠊࢩࣕࢪࢡࣔ⸴㢮㝣ୖ᳜≀ࡢࢤࣀ࣒ࣞ࣋ࣝ ࡢ㐪࠸ࡢእほࡀぢ࠼࡚ࡁࡓࠋࡲࡓࠊಶࠎࡢ㑇ఏᏊࡢ ゎᯒࡼࡾࠊࡑࡢ♽ඛⓗ࡞ᛶ㉁㐍㐣⛬ࢆ᥎ᐃࡍ ࡿࡇࡀ࡛ࡁࡓࠋᚋࠊࢩࣕࢪࢡࣔࡢⓎ⏕㐣⛬࠾ ࡅࡿ⥙⨶ⓗ࡞㑇ఏᏊⓎ⌧ゎᯒࡸಶࠎࡢ㑇ఏᏊࡢᶵ⬟ ゎᯒࡀ㐍ᒎࡋࠊ⣔⤫㛫࡛ࡢẚ㍑ࡀྍ⬟࡞ࡿࡇ ࡼࡾࠊࢩࣕࢪࢡࣔ⸴㢮㝣ୖ᳜≀࠾ࡅࡿከ⣽⬊య ไࡢ㉳※㐍㐣⛬ࡀ᫂☜࡞ࡗ࡚ࡃࡿᮇᚅࡉࢀ ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࢩࣕࢪࢡࣔࡘ࠸࡚ࠊᮏᨭࢆཷࡅࡓඹྠ◊✲㸦ᨭ ౫㢗⪅㸸㔠ἑ࣭すᒣᬛ᫂༤ኈ㸧ࡼࡾ᫂ࡽ ࡋࡓᴫせࢤࣀ࣒ࢆࡶࠊࢩࣕࢪࢡࣔࡢⓎ⏕ẁ㝵ࢆ ࡰ⥙⨶ࡍࡿ RNA-seq ࢹ࣮ࢱࠊ࠾ࡼࡧ㝣ୖ᳜≀ࡢࢱ ࣥࣃࢡ㉁ࡢ࣑ࣀ㓟㓄ิࢆ⏝࠸㑇ఏᏊࣔࢹࣝᵓ⠏ࢆ ⾜࠸ࠊ35,883 㑇ఏᏊࠊ36,887 ㌿⏘≀ࢆண ࡋࡓࠋ ࢩࣕࢪࢡࣔຍ࠼ࠊࢥࣞ࢜ࢣ࣮ࢸ㢮ࠊ࣍ࢩ࣑ࢻࣟ㢮ࠊ ࢡࣞࣈࢯ࣑ࣝࢹ࣒࢘ࠊࢡ࣑ࣛࢻࣔࢼࢫࡢࢤࣀ࣒/ࢺ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Sato, M. et al. 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Phycol. 47, 999–1008 (2011) - 355 - බເᨭㄢ㢟㸸2013 ᖺᗘ ከᩘࡢ ⣽⬊ࡽᐃ㔞ᛶࡢ࠶ࡿ⥙⨶ⓗ㑇ఏᏊⓎ⌧ሗࢆྲྀࡾฟࡍࡓࡵࡢᇶ┙ᢏ⾡ࡢ㛤Ⓨ ᣮᡓⓗⴌⱆ◊✲ ➲ᕝ ὒᖹ࣭⌮Ꮫ◊✲ᡤሗᇶ┙ࢭࣥࢱ࣮ࣂ࢜ࣥࣇ࢛࣐ࢸࢡࢫ◊✲㛤Ⓨࣘࢽࢵࢺ 㸺⫼ᬒ࣭┠ⓗ㸼 ᵝࠎ࡞⏕⌮᮲௳࠾࠸࡚㞟ᅋ⣽⬊ෆ࡛ࡣࠊࢤࣀ࣒ ሗࡀྠ୍࡛࠶ࡗ࡚ࡶࠊ ⣽⬊㛫 51$ ࡸࢱࣥࣃࢡ㉁ࡢ Ⓨ⌧㔞ࡢᆒ୍ᛶࡀᏑᅾࡍࡿࠋᇵ㣴ࡋࡓࡼࡾᆒ୍࡞ ⣽⬊㞟ᅋ࠾࠸࡚ࡶࠊ51$ ࣞ࣋ࣝࡢⓎ⌧㔞ࡢࡤࡽࡘࡁ ࡀᏑᅾࡋࠊ⣽⬊ᶵ⬟ࡢ㛵㐃ᛶࡶ♧၀ࡉࢀ࡚࠸ࡿࠋ ཎ⌮ⓗࠊ ⣽⬊ 51$VHT ἲࡣ ⣽⬊㛫ࡢ 51$ ࣞ࣋ࣝ ࡢⓎ⌧㔞ࡢኚືࢆ⥙⨶ⓗ᳨ฟ࡛ࡁࡿྍ⬟ᛶࢆࡶࡘࠋ ࡋࡋࠊࡼࡾᚤ⣽࡞㑇ఏᏊⓎ⌧ࡢࡤࡽࡘࡁࢆ᳨ฟࡍ ࡿࡣᐃ㔞ᛶ㸦᳨ฟ㑇ఏᏊᩘ࣭⌧ᛶࡸṇ☜ᛶ㸧ࡢ ㊊ࡀᣦࡉࢀ࡚࠸ࡓࠋࡇࢀࡲ࡛ᡃࠎࡣ⡆౽࡛ᐃ 㔞ᛶࡢ㧗࠸ ⣽⬊ 51$VHT㸦4XDUW]6HT㸧ἲࢆ㛤Ⓨࡋࠊ 㑇ఏᏊⓎ⌧ࡢࡤࡽࡘࡁࢆ⥙⨶ⓗㄪࡿࡇᡂຌ ࡋ࡚࠸ࡿࠋ୍᪉࡛⌮ⓗ࡞ ⣽⬊ 51$VHT ࡢࡓࡵ ࡣࠊᐃ㔞ᛶࡢࡉࡽ࡞ࡿᨵၿࡸࠊ୍ᗘฎ⌮ྍ⬟࡞ࢧ ࣥࣉࣝᩘࡢୖ᪼㸦ࣁࢫ࣮ࣝࣉࢵࢺᛶ⬟㸧ࡸ QRQSRO\$51$ ࡢ᳨ฟࡀồࡵࡽࢀࡿࠋࡇࢀࡽࢆࡍ ࡚‶ࡓࡍ᪉ἲࡣࠊ4XDUW]6HT ࢆྵࡵ୍ࡘࡶ࡞ࡃࠊヱ ᙜ᪉ἲࡢ㛤Ⓨࢆヨࡳࡓࠋ 㸺ᨭࡢෆᐜ㸼 4XDUW]6HT ࢆྵࡵࡓሗ࿌ࡉࢀ࡚࠸ࡿࡰ࡚ࡢ ⣽ ⬊ 51$VHT ἲࡣࠊROLJRG7 ࣉ࣐࣮ࣛࢆά⏝ࡍࡿࡇ ࡛ SRO\$51$ ᳨ฟᑐ㇟ࡋࠊ̓ᮎ➃ࡽ ̓ᮎ➃ ࡅࡿࡲ࡛ࢆ ༢ࡍࡿẚ㍑ⓗ㛗࠸ F'1$ ࡋ࡚ '1$ ቑᖜࢆ⾜ࡗ࡚࠸ࡓࠋࡑࡢࡓࡵቑᖜࣂࢫࡀ ࡾࡸࡍࡃࠊ4XDUW] ࠾ࡅࡿቑᖜ࡛ࡣࣂࢫࡀ ࡽ࡞࠸ࡼ࠺ࡃ 3&5 ቑᖜࢆᨵၿࡋ࡚ࡁࡓࡀࠊ᳨ ฟឤᗘࡢᨵၿࡣࡇࢀ௨ୖ㞴ࡋ࠸ࠋ୍᪉࡛ࠊࣛࣥࢲ࣒ ࣉ࣐࣮ࣛ࡞࡛㏫㌿ࡍࡿ ̓ᮎ➃ࡽ ̓ᮎ➃ ࡅࡿࡲ࡛ࢆᩘ༑༢ศ࡛ࡁࡿࡓࡵࠊቑᖜࣂ ࢫࢆࣜࢫࢡศᩓ࡛ࡁࡿྍ⬟ᛶࡀ࠶ࡿࡓࡵ᳨ฟឤ ᗘࡀୖ᪼ࡍࡿࡇࡀᮇᚅࡉࢀࡿࠋᑐ㇟ 51$ ࡢ SRO\$ 㒊ศࢆ͇Ⅼ͇࡛ᤊ࠼ࡿ᪉ᘧࡽࠊᑐ㇟ 51$ ࢆศࡋ ࡚͇㠃͇࡛ᤊ࠼ࡿ᪉ᘧࡍࡿࡇ࡛ࠊ᳨ฟឤᗘࢆୖ ᪼ࡉࡏࡿࡇࢆヨࡳࡓࠋ᭱ึࣛࣥࢲ࣒ࣉ࣐࣮ࣛ ࣮࣋ࢫࡢ㏫㌿ᛂࡢ᭱㐺ࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊ ᪤Ꮡ᪉ἲ࡛ࡣࠊᩘ FRS\ ࡢ㑇ఏᏊࢆᤊ࠼ࡿࡇࡀ㞴ࡋ ࡗࡓࡀࠊᨵၿ᪉ἲ࡛ࡣࡰ ᤊ࠼ࡿࡇࡀฟ᮶ ࡓࠋࡇࢀࢆⓎᒎࡉࡏࠊ㉸୪ิᆺࢩ࣮ࢣࣥࢧ࣮ࡢࣛ ࣈࣛࣜస〇ࢆヨࡳࠊ࠸ࡃࡘࡢ᪉ᘧࡢ ⣽⬊ 51$VHT ࡢࢩ࣮ࢣࣥࢫࣛࣈࣛࣜ '1$ ࢆస〇ࡋࡓࠋ ࢤࣀ࣒ᨭ࡛ࡣࠊᮾிᏛ㕥ᮌ✨ඛ⏕㕥 ᮌ◊✲ᐊࡢⓏᆏᵝࠊᇼෆᵝࠊ⳥ụᵝᨭ࠸ࡓࡔ࠸ ࡓࠋ㛤Ⓨἲ࡛స〇ࡋࡓࢩ࣮ࢣࣥࢫࣛࣈࣛࣜ '1$ ࢆ ,OOXPLQD+L6HT3DLUHGHQGES ࡛ ࣮ࣞࣥศゎ ᯒࡋ࡚࠸ࡓࡔ࠸ࡓࠋ⥲ᩘ ࢧࣥࣉࣝࠊ ⣽⬊ 51$VHT ἲ ✀㢮ࡢゎᯒࢆࡋ࡚࠸ࡓࡔࡁࠊ୍㒊ࢩ࣮ࢣࣥࢫࢹ ࣮ࢱࡀ㊊ࡋࡓ㒊ศ㛵ࡋ࡚ࡣࠊ㏣ຍࡢࢩ࣮ࢣࣥࢫ ࡶ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 >⡆౽࡛ࡘࠊ㧗࠸ᐃ㔞ᛶ⬟ࢆⓎࡍࡿࠊQRQSRO\$ 51$ ࢆ᳨ฟ࡛ࡁࡿ㛤Ⓨ᪉ἲ 6&3@ PRXVH(6 ⣽⬊⏤᮶ SJWRWDO51$ ࢆேᕤࢧࣥࣉࣝ ࡋ࡚⏝࠸ࡓᛶ⬟ẚ㍑ࡢ⤖ᯝࠊ࢜ࣜࢪࢼࣝ㛤Ⓨἲ 6&3 ࡀࡶࡗࡶ㧗࠸ᐃ㔞ᛶ⬟ࢆⓎࡍࡿࡇࡀࢃ ࡗࡓࠋXJ ࡢ WRWDO51$ ࢆ⏝࠸ࡓ㏻ᖖࡢ 51$VHT ࡢ ࢹ࣮ࢱᕼ㔘ࡼࡾస〇ࡋࡓ SJWRWDO51$ ࢆ⏝ ࠸ࡓ ⣽⬊ 51$VHT ࡢࢹ࣮ࢱࢆẚ㍑ࡍࡿࡇ࡛ࠊ ࢀࡃࡽ࠸ࡶࡢࣉࣟࣇࣝࢆಖࡗ࡚࠸ࡿㄪࡿ ࡇࡀ࡛ࡁࡿࠋ4XDUW]6HT ࡛ࡣࠊ ࢺࣛࣥࢫࢡ ࣜࣉࢺࡀ᳨ฟࡉࢀ ಸࡢ⠊ᅖෆࡲࡿࢺࣛࣥࢫࢡ ࣜࣉࢺࡀ⣙ ࡢ 㑇ఏᏊࡀ᳨ฟࡉࢀࡓࠋ୍᪉࡛ࠊ 6&3 ࡛ࡣ⣙ ࢺࣛࣥࢫࢡࣜࣉࢺࡀ᳨ฟࡉࢀࠊ ಸࡢ⠊ᅖෆ⣙ ࢺࣛࣥࢫࢡࣜࣉࢺࡀ᳨ฟ ࡉࢀࡓࠋ6&3 ྠᵝࣛࣥࢲ࣒ࣉ࣐࣮ࣛࢆ⏝࠸ࡓ ᕷ㈍ရࡢሙྜࠊ ࢺࣛࣥࢫࢡࣜࣉࢺࡀ᳨ฟࡉࢀࠊ ⣙ ࡢ ࢺࣛࣥࢫࢡࣜࣉࢺࡀ ಸࡢ⠊ᅖෆ᳨ ฟࡉࢀࡓࠋࡇࢀࡽࡢ⤖ᯝࡽࠊ㛤Ⓨࡋࡓ᪉ἲࡀࠊ ⣽⬊ࡽ᭱ࡶᐃ㔞ᛶⰋࡃ㑇ఏᏊⓎ⌧㔞ࢆྲྀࡾฟࡏࡿ ࡇࡀࢃࡗࡓࠋࡲࡓࠊ6&3 ࡛ࡣࠊ+LVWKE ࡞ࡢ SRO\$ ࢸ࣮ࣝࢆᣢࡓ࡞࠸ࢺࣛࣥࢫࢡࣜࣉࢺࡀ᳨ฟ࡛ ࡁࡓࠋ4XDUW]6HT ࡞ࡢ ⣽⬊ SRO\$51$VHT ἲ࡛ ฟ᮶࡞ࡗࡓ QRQSRO\$51$ ࡢ᳨ฟᡂຌࡋࡓࠋ㛗 㙐ࢺࣛࣥࢫࢡࣜࣉࢺ࠾ࡅࡿ㛗ࡢ࢝ࣂࣞࢵࢪࡢᨵ ၿࡶ☜ㄆ࡛ࡁࡓࠋ6&3 ࡛ࡣ 4XDUW]6HT ࡃࡽ࡚ࢫ ࢸࢵࣉᩘࡀ༙ศ࡞ࡾ⡆౽ᛶࡀࡉࡽᨵၿࡋࡓࠋ୍ ᪉࡛ࠊU51$ ࡀయࡢ࣮ࣜࢻࡢ ⛬ࢆࡋࡵࡿࡇࡀ ࢃࡗࡓࠋ࣑ࢺࢥࣥࢻࣜ⏤᮶ 51$ ࡞ࢆྵࡵࠊᨵ ၿࡢవᆅࡀ࠶ࡿࡇࡶศࡗࡓࠋ ṧࡿࢧࣥࣉࣝࡢゎᯒࡶࡍࡍࡵࠊQRQSRO\$ 51$ ࢆ᳨ฟྍ⬟࡞᪂つ ⣽⬊ 51$VHT ἲࡋ࡚ࠊㄽᩥ ࢆ㐍ࡵࡿணᐃ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ⛉Ꮫ◊✲㈝ࠕᣮᡓⓗⴌⱆ◊✲ࠖࡢ⠊ᅖෆ࡛ࡣ㞴ࡋ࠸ つᶍࡢࢩ࣮ࢣࣥࢫ㔞㸦51$VHT ࢧࣥࣉࣝ㸧ࢆࡋ࡚ ࠸ࡓࡔࡃࡇ࡛ㄢ㢟ࢆࡁࡃᣑࡉࡏࡿࡇࡀྍ⬟ ࡞ࡗࡓࠋᡃࠎࡢ㛤Ⓨࡋࡓ͆ᩘ FRS\ ࢆᤊ࠼₃ࡽࡉ࡞ ࠸᪂つ㏫㌿ᛂ㸦≉チ⏦ㄳ୰㸧͇ࢆ ⣽⬊ 51$VHT ἲᛂ⏝ࡋࠊ㠀ᖖ㧗࠸ຠᯝᐇ⏝ᛶࢆ♧ࡍࡇࡀ ࡛ࡁࡓࠋࡇࢀࡲ࡛࡛ࡁ࡞ࡗࡓ QRQSRO\$51$ ࡢ᳨ ฟࡀྍ⬟࡞ࡾࠊ ⣽⬊࡛ࡢ QRQSRO\$ ࢺࣛࣥࢫࢡ ࣜࣉࢺゎᯒࡢ㐨ࡀᣅࡅࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 356 - බເᨭㄢ㢟㸸2010 ᖺᗘ ᶆⓗ㑇ఏᏊࣀࢵࢡࢲ࢘ࣥࡼࡿ㟋㛗㢮ࣄࢺᝈࣔࢹࣝࡢసฟ ᇶ┙◊✲㸦㸿㸧 బࠎᮌ ࠼ࡾ࣭ᐇ㦂ື≀୰ኸ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢥ࣐࣮ࣔࣥࣔࢭࢵࢺ㸦࣐࣮ࣔࢭࢵࢺ㸧ࡣᑠᆺࡢ㟋 㛗㢮࡛࠶ࡾࠊ⦾Ṫᛶࡀ㧗ࡃࠊே⋇ඹ㏻ឤᰁࢆᣢࡓ ࡞࠸ࠊḟୡ௦ᑟධ㑇ఏᏊࡀఏ㐩ྍ⬟࡞㑇ఏᏊᨵ ኚື≀ࡢసฟࡀྍ⬟࡞࡞ࡽࣂ࣓࢜ࢹ࢝ࣝ ࢧ࢚ࣥࢫࡢᐇ㦂ື≀ࡋ࡚᭷⏝࡛࠶ࡿࠋ ᮏ◊✲࡛ࡣࠊⓎ⌧㑇ఏᏊࢆศゎࡍࡿ RNA ᖸ΅⌧ ㇟ࢆᛂ⏝ࡋࠊ┠ⓗࡢ㑇ఏᏊࡢࡳࢆᢚไⓗࡘྍ㏫ⓗ ไᚚࡍࡿ㑇ఏᏊࣀࢵࢡࢲ࢘ࣥ㸦KD㸧࣐࣮ࣔࢭࢵࢺ ࡢసฟἲࢆ☜❧ࡋࠊᚓࡽࢀࡓ KD ࣐࣮ࣔࢭࢵࢺࡢࣄ ࢺᝈࣔࢹࣝື≀ࡋ࡚ࡢ᭷ຠᛶࢆ᫂ࡽࡍࡿ ࢆ┠ⓗࡍࡿࠋࡋࡋ࡞ࡀࡽࠊࡇࢀࡽࣀࢵࢡ࢘ࢺ ࣐࣮ࣔࢭࢵࢺࠊࣀࢵࢡࢲ࣐࣮࢘ࣥࣔࢭࢵࢺࡢసฟ ࡣࢤࣀ࣒ࢡ࣮ࣟࣥࡀᚲせ࡛࠶ࡿࠋ⌧ᅾࠊ࣐࣮ࣔࢭࢵ ࢺࢤࣀ࣒ BAC ࣛࣈ࣮ࣛࣜࡣసᡂࡉࢀ࡚࠸ࡿࡀࠊᚲ せࡍࡿࢡ࣮ࣟࣥࡢ㑅ᢥࡣࠊࣇࣝࢱ࣮ࣁࣈࣜ ࢲࢮ࣮ࢩࣙࣥࠊPCR ࡞ࡢࢫࢡ࣮ࣜࢽࣥࢢࡀᚲせ ࡛࠶ࡾࠊࣀࢵࢡ࢘ࢺࠊࣀࢵࢡࢲ࢘ࣥ࣋ࢡࢱ࣮ࢆస ᡂࡍࡿࡣከ࡞㛫ປຊࡀࡗ࡚࠸ࡿࠋࡑࡇ ࡛࣐࣮ࣔࢭࢵࢺ BAC ࢡ࣮ࣟࣥࡢ㓄ิሗࢆࢤࣀ࣒ ୖ࣐ࢵࣉࡍࢀࡤࠊ↹㞧࡞ࢫࢡ࣮ࣜࢽࣥࢢ㐣⛬ࢆ⤒ ࡿࡇ࡞ࡃࢥࣥࣆ࣮ࣗࢱୖ࡛ᕼᮃࡍࡿࢡ࣮ࣟࣥࢆ᳨ ⣴࣭ྠᐃࡍࡿࡇࡀྍ⬟࡛࠶ࡿࠋᚋࠊከࡃࡢ㑇ఏ Ꮚᨵኚ࣐࣮ࣔࢭࢵࢺࡢసฟࡀぢ㎸ࡲࢀࠊ㟋㛗㢮ࡢᐇ 㦂ື≀ࡋ࡚⏝ቑຍࡀぢ㎸ࡲࢀ࡚࠸ࡿ࣐࣮ࣔࢭࢵ ࢺࡢ BAC ࣛࣈ࣮ࣛࣜࡣᅜࡋ࡚ᡓ␎ⓗᩚഛࡍ ࡁࣜࢯ࣮ࢫ࡛࠶ࡾࠊࡑࡇ㓄ิሗࢆຍࡍࡿࡇ ࡼࡾࠊࡇࢀࡽࡢࢡ࣮ࣟࣥࢆ⏝࠸ࡓᶵ⬟ࢤࣀ࣒ゎ ᯒࠊࡦ࠸࡚ࡣᅜෆእࡢࣛࣇࢧ࢚ࣥࢫ◊✲ࡀ㣕㌍ ⓗⓎᒎࡍࡿ⪃࠼ࡽࢀࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࣐࣮ࣔࢭࢵࢺ ES ⣽⬊ࡼࡾศㄏᑟࡋࡓ⥺⥔ⱆ⣽⬊ ᰴࢆㄪ〇ࡋࠊࡇࡢ⣽⬊ࡽᚓࡽࢀࡓ࣐࣮ࣔࢭࢵࢺࢤ ࣀ࣒ DNA ࢆ⏝࠸࡚సᡂࡋࡓ ES ⏤᮶ࡢࣛࣈ࣮ࣛࣜ (384 well plate, 220 ᯛศ㸧ࡢ࢚ࣥࢻࢩ࣮ࢡ࢚ࣥࢫࢆ ⾜ࡗ࡚࠸ࡓࡔࡁࠊ࡚ࡢ࢚ࣥࢻࢩ࣮ࢡ࢚ࣥࢫࡀ⤊ ࡋࡓࠋ ඹྠࡋ࡚ࠊBAC ࣛࣈ࣮ࣛࣜࡢ࢚ࣥࢻࢩ࣮ࢡ࢚ࣥࢫ 㓄ิࢆࠊᐇ୰◊࣐࣮ࣔࢭࢵࢺࢤࣀ࣒㓄ิୖ BLAT ࢆ⏝࠸࡚࣐ࢵࣆࣥࢢࢆ⾜࠸ࠊBAC ࣛࣈ࣮ࣛࣜ≀⌮ ᆅᅗࢆసᡂࡋࡓࠋḟࠊࡇࡢ BAC ᆅᅗࢆ࣐࣮ࣔࢭࢵ ࢺࢤࣀ࣒ࡢ⤫ྜࣈࣛ࢘ࢨ࣮ୖᐇࡋࠊBAC ࣛࣈ ࣮ࣛࣜࢆ⏝࠸ࡓ㑇ఏᏊᕤᏛᐇ㦂ࢆ⾜࠺࣐࣮ࣔࢭࢵࢺ ◊✲⪅බ㛤ࡋࡓࠋBAC ࣛࣈ࣮ࣛࣜࢆ⏝࠸ࡓ࣐࣮ ࣔࢭࢵࢺ㑇ఏᏊᨵኚᐇ㦂᭷ຠ⏝ࡍࡿࡇࡼࡾࠊ ᙜヱ◊✲ศ㔝ࡀຍ㏿ࡉࢀࡿࡇࡀᮇᚅࡉࢀࡿࠋࡉࡽ ࠊBAC ࣛࣈ࣮ࣛࣜࡢᡂᯝࡶྵࡵࡓㄽᩥࢆⓎ⾲ࡋ ࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ᮇ㛫㑇ఏᏊᨵኚᢏ⾡ࢆྲྀࡾᕳࡃ⎔ቃࡀࡁ ࡃኚࡋࠊࢤࣀ࣒⦅㞟ᢏ⾡࠸࠺᪂ࡓ࡞ᢏ⾡ࡀฟ⌧ ࡋࠊ࣓࢟ࣛಶయస〇⬟ࢆᣢࡘ ES ⣽⬊ࠊiPS ⣽⬊ࢆస 〇࡛ࡁ࡞࠸ື≀✀࡛ࡶᶆⓗ㑇ఏᏊࣀࢵࢡ࢘ࢺಶయ ࡢస〇ࡀẚ㍑ⓗᐜ᫆ྍ⬟࡞ࡗࡓࠋࡋࡋ࡞ࡀࡽࠊ ᶆⓗ㑇ఏᏊࣀࢵࢡࣥᢏ⾡ࡣࠊࡑࡢస〇ຠ⋡ࡽ㠀 ࣄࢺ㟋㛗㢮࡛ࡣ⌧ᅾࡢࢤࣀ࣒⦅㞟ᢏ⾡࡛ࡣᮍࡔᅔ㞴 ࡛࠶ࡿࠋᮏᨭࡼࡗ࡚ BAC ࣛࣈ࣮ࣛࣜࡢᩚഛࡀ ᩚ࠸ࠊᑐ㇟ࡢ㑇ఏᏊࡢ㏆ഐ㡿ᇦࡢࣉ࣮ࣟࣔࢱࠊ࢚ࣥ ࣁࣥࢧ࣮ࡢゎᯒࢆ⾜ࢃ࡞ࡃ࡚ࡶṇ☜࡞⤌⧊≉␗ⓗⓎ ⌧ࡍࡿ㑇ఏᏊᨵኚ࣐࣮ࣔࢭࢵࢺࢆస〇ࡍࡿࡇࡀྍ ⬟࡞ࡗࡓࠋ㏆ᖺࠊࡆࡗṑ㢮㟋㛗㢮࡛ࡣࠊᵝࠎ࡞ ⤌⧊࡛Ⓨ⌧ࡍࡿ㑇ఏᏊࡀ␗࡞ࡿࡇࠊࡲࡓࡣⓎ⌧ ࣃࢱ࣮ࣥࡀ␗࡞ࡿࡇࡀ᫂ࡉࢀ࡚ࡁ࡚࠸ࡿࡀࠊ 㟋㛗㢮࠾ࡅࡿࡇࢀࡽࡢ㑇ఏᏊᶵ⬟ࡢゎᯒࡀྍ⬟ ࡞ࡗࡓࠋᚋࡣࠊ㟋㛗㢮≉᭷࡛࠶ࡿ㧗ḟㄆ▱ ᶵ⬟ࡀࡢࡼ࠺Ⓨ㐩ࡍࡿࡢ࠸࠺ၥ㢟ࢆ᫂ ࡋ࡚࠸ࡃୖ࡛㔜せ࡞ᇶ┙࡞ࡿࡶࡢ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Sato K et al., Scientific Reports, 5, 2015, DOI: 16894-16901 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᠕⩏ሿᏛ⌮ᕤᏛ㒊⏕ሗ⛉Ꮫ⛉ᴬཎ◊✲ᐊ - 357 - බເᨭㄢ㢟㸸2014 ᖺᗘ Hippo ࢩࢢࢼࣝ⤒㊰ࡼࡿ⣽⬊㛫ࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡢศᏊᇶ┙ ᇶ┙◊✲㸦㸿㸧 బࠎᮌ ὒ࣭⇃ᮏᏛⓎ⏕་Ꮫ◊✲ᡤ බເᨭㄢ㢟㸸2015 ᖺᗘ ⥺⥔ⱆ⣽⬊࠾ࡅࡿ⣽⬊➇ྜᶵᵓࡑࡢⓎ⏕Ꮫⓗព⩏ ᪂Ꮫ⾡㡿ᇦ◊✲ బࠎᮌ ὒ࣭㜰ᏛᏛ㝔⏕ᶵ⬟◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⚾ࡓࡕࡢయࡣࠊࡓࡗࡓ୍ࡘࡢཷ⢭༸ࡽࠊⓎ⏕ ࡼࡗ࡚ṇ☜⌧ᛶⰋࡃసࡽࢀࡿࠋⓎ⏕ࡢ㔜せ࡞≉ ᚩࡢ୍ࡘࡣࡑࡢṇ☜ᛶ࡛࠶ࡿࡀࠊ⬇ෆࡢಶࠎࡢ⣽⬊ ࡢᣲືࡣࡤࡽࡘࡁࡀ࠶ࡾࠊ࡞ࡤࡽࡘࡁࢆᣢࡗࡓື ࡁࢆࡍࡿ⣽⬊ࡀࠊ㞟ᅋࡋ࡚ṇ☜⌧ᛶⰋࡃయࢆ ࡘࡃࡾୖࡆࡿࡓࡵࡣࠊ㞄᥋ࡍࡿ⣽⬊ྠኈࡀࢥ࣑ࣗ ࢽࢣ࣮ࢩࣙࣥࡍࡿࡇ࡛ࠊࡑࡢᣲືࢆㄪࡉࡏࡿࡇ ࡀᚲせ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋࡋࡋࠊࡑࡢࡼ࠺࡞ࠊ 㞄᥋⣽⬊ࡢ≧ែࢆㄆ㆑ࡋࡓ⣽⬊㛫ࢥ࣑ࣗࢽࢣ࣮ࢩࣙ ࣥࡢ⤌ࡳࡣ࠶ࡲࡾࢃࡗ࡚࠸࡞࠸ࠋ ⚾ࡣࡇࢀࡲ࡛ࡢ࣐࢘ࢫ⬇Ⓨ⏕ࡢ◊✲ࡼࡗ࡚ࠊ Hippo ࢩࢢࢼࣝ⤒㊰ࡀ⣽⬊㛫ࡢ᥋╔ࡼࡗ࡚άᛶ ࡉࢀࡿࢩࢢࢼ࡛ࣝ࠶ࡾࠊ⣽⬊ࡀࠊ⣽⬊ᐦᗘ࡞ࢆㄆ ㆑ࡍࡿ㛵ࢃࡗ࡚࠸ࡿࡇࢆぢฟࡋࡓࠋࡉࡽࠊ ࣐࢘ࢫ⬇⥺⥔ⱆ⣽⬊⏤᮶ࡢ NIH3T3 ⣽⬊࠾࠸࡚ࠊ Hippo ⤒㊰ࡢୗὶ࡛ാࡃ㌿ᅉᏊ Tead ࡢ㌿άᛶ ࡢ␗࡞ࡿ⣽⬊ࢆΰࡐ࡚ඹᇵ㣴ࡍࡿࠊTead άᛶࡢ㧗 ࠸⣽⬊ࡀඃඛⓗቑṪࡋࠊTead άᛶࡢప࠸⣽⬊ࡀ 㝖ࡉࢀࡿ⣽⬊㛫ࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡀ㉳ࡇࡿࡇࢆ ぢฟࡋࡓࠋࡇࡢࡼ࠺࡞≧ែ(㐺ᛂᗘ࠸࠺)ࡢ␗࡞ࡿ㞄 ᥋⣽⬊㛫ぢࡽࢀࡿ⣽⬊㛫ࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡣࠊ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚࡛᭱ึྠᐃࡉࢀࡓ⣽⬊➇ྜ ࡤࢀࡿࡶࡢ࡛࠶ࡿࠋ ⣽⬊➇ྜࡢ◊✲ࡣࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ࢆ୰ᚰ㐍ࡵ ࡽࢀ࡚࠸ࡿࡀࠊࢩࣙ࢘ࢪࣙ࢘ࣂ࢚࠾࠸࡚ࡶࠊࡢ ࡼ࠺ࡋ࡚㞄᥋⣽⬊ࡢ≧ែࢆㄆ㆑ࡍࡿࡢ࡞ࠊࡑ ࡢࡋࡃࡳࡢከࡃࡀࢃࡗ࡚࠸࡞࠸ࠋ့ங㢮࠾࠸࡚ ࡣࠊ࠸ࡃࡘࡢ⣔࡛⣽⬊➇ྜࡀ㉳ࡇࡿሗ࿌ࡀ࠶ࡿࡶ ࡢࡢࠊࡑࡢࡋࡃࡳࡸⓎ⏕࠾ࡅࡿᙺ࡞ࡣࠊ ࢇࢃࡗ࡚࠸࡞࠸ࡢࡀ⌧≧࡛࠶ࡿࠋ ᮏ◊✲࡛ࡣࠊ࣐࢘ࢫ࠾ࡅࡿ㞄᥋⣽⬊ࡢ≧ែࢆㄆ ㆑ࡋࡓ⣽⬊㛫ࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࡢ⤌ࡳࢆ᫂ࡽ ࡍࡿࡇࢆ┠ⓗࡋ࡚ࠊᡃࠎࡀᶞ❧ࡋࡓ NIH3T3 ⣽⬊ࢆ⏝࠸ࡓ⣽⬊➇ྜࡢ⣔ࢆ⏝ࡋ࡚ࠊ⣽⬊➇ྜ 㛵ࢃࡿ㑇ఏᏊࡢ⥙⨶ⓗ࡞ࢫࢡ࣮ࣜࢽࣥࢢࢆ࠾ࡇ࡞ࡗ ࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࢫࢡ࣮ࣜࢽࣥࢢࡢ⣔ࡋ࡚ࠊ௨ୗࡢࡼ࠺࡞᪉ἲࢆ ⏝࠸ࡓࠋࡲࡎࠊCRISPR/Cas9 ࢩࢫࢸ࣒ࡢ gRNA ࡢ ࢤࣀ࣒࣡ࢻ࡞ࣞࣥࢳ࢘ࣝࢫࣛࣈ࣮ࣛࣜࢆࠊ Cas9 ࢆⓎ⌧ࡉࡏࡓ NIH3T3 ⣽⬊ឤᰁࡉࡏࡿࡇ ࡛ࠊ ᵝࠎ࡞㑇ఏᏊኚ␗ࢆᣢࡘ NIH3T3 ⣽⬊ࡢ㞟ᅋࠊ ࡍ࡞ࢃࡕኚ␗⣽⬊ࣛࣈ࣮ࣛࣜࢆసᡂࡍࡿࠋࡇࡢኚ ␗⣽⬊ࣛࣈ࣮ࣛࣜࢆᇵ㣴ࡍࡿࠊ⣽⬊➇ྜ㛵ࢃ ࡿ㑇ఏᏊኚ␗ࢆᣢࡘ⣽⬊ࡣࠊ㛫ࡶࠊࡑࡢ Ꮡᅾẚ⋡ࡀኚࡍࡿ⪃࠼ࡽࢀࡿࠋࡍ࡞ࢃࡕࠊ⣽⬊ ➇ྜࡢ⪅࡞ࡿ⣽⬊ࡣࢥࣥࣇ࢚ࣝࣥࢺ࡞ࡗ࡚ࡶ ⣽⬊ቑṪࢆ⥆ࡅࡿࡓࡵᏑᅾẚ⋡ࡀቑຍࡍࡿࠋ୍᪉ࠊ ᩋ⪅࡞ࡿ⣽⬊ࡣቑṪࡀᢚไࡉࢀ㝖ࡉࢀࡿࡓࡵࠊ 㛫ඹᏑᅾẚ⋡ࡀῶᑡࡍࡿࠋࡑࡇ࡛ࠊඖࡢ gRNA ࣛࣈ࣮ࣛࣜࠊᇵ㣴 4 ᪥┠ 10 ᪥┠ࡢኚ␗⣽⬊ࣛ ࣈ࣮ࣛࣜ࠾ࡅࡿྛ gRNA ࡢᏑᅾẚ⋡ࢆẚ㍑ࡋࠊᏑ ᅾẚ⋡ࡢ⤒ⓗ࡞ቑῶࡋ࡚ࠊ⣽⬊➇ྜ㛵㐃㑇ఏᏊ ࢆྠᐃࡍࡿࠋ ᮏᨭ࡛ࡣࠊඖࡢ gRNA ࣛࣈ࣮ࣛࣜࠊᇵ㣴 4 ᪥ ┠ 10 ᪥┠ࡢኚ␗⣽⬊ࣛࣈ࣮ࣛࣜࡢࢤࣀ࣒ DNA ྵࡲࢀࡿ gRNA ࢆḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮࡛ゎㄞࡋࠊ ࡉࡽࠊྛ gRNA ࡢᏑᅾẚ⋡ࡢ⤒ⓗ࡞ኚࢆゎᯒ ࡋ࡚࠸ࡓࡔ࠸ࡓࠋ2014 ᖺᗘࡣ Koike-Yusa ࡽࡼࡿ gRNA ࣛ ࣈ ࣛ ࣜ ࣮ (Nat Biotech 32, 267-273, 2014)ࢆ⏝࠸ࡓࢫࢡ࣮ࣜࢽࣥࢢࢆ 2 ᅇ⾜࠸ࠊ2015 ᖺ ᗘࡣࠊࡇࢀࡲ࡛ Zhang ࡽࡼࡿ gRNA ࣛࣈࣛࣜ ࣮(Nat Methods 11, 783-784, 2014)ࢆ⏝࠸ࡓࢫࢡࣜ ࣮ࢽࣥࢢࢆ 2 ᅇ࠾ࡇ࡞ࡗࡓࠋ ḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮ࡼࡿሷᇶ㓄ิࡢゎㄞࡣᅜ ❧㑇ఏᏛ◊✲ᡤࡢ㇏⏣ᩔඛ⏕ࠊࢹ࣮ࢱࡢゎᯒࡣᮾ ிᏛࡢᒾᓮ΅ඛ⏕ࠊᯇồඛ⏕ࠊࡑࢀࡒࢀᨭ ࡋ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 <2014 ᖺᗘ㸼 2014 ᖺᗘࡣ Koike-Yusa ࡽࡢ gRNA ࣛࣈ࣮ࣛࣜ (Nat Biotech 32, 267-273, 2014)ࢆ⏝࠸࡚ NIH3T3 ࡢኚ␗⣽⬊ࣛࣈ࣮ࣛࣜసᡂࡋࠊࢫࢡ࣮ࣜࢽࣥࢢࢆ ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊᮏ᪉ἲࡼࡗ࡚ࠊᐇ㝿⣽⬊➇ ྜ㛵㐃ࡍࡿ㑇ఏᏊࡢྠᐃࡀྍ⬟࡛࠶ࡿࡇࡀ☜ㄆ ࡛ࡁࡓࠋࡲࡓࠊࡑࡢ୍᪉࡛ࠊࡣࡗࡁࡾࡋࡓุᐃࡀ ࡛ࡁ࡞࠸㑇ఏᏊࡶከࡃᚓࡽࢀࠊࡉࡽ࡞ࡿ᪉ἲࡢᨵၿ ࡀᚲせ࡛࠶ࡿࡇࡶ᫂ࡽ࡞ࡗࡓࠋලయⓗࡣࠊ ௨ୗࡢ࠾ࡾ࡛࠶ࡿࠋ Hippo ⤒㊰␗ᖖࢆᣢࡘ⣽⬊ࡣࠊ⣽⬊➇ྜࡢ⪅ ࡞ࡾࠊ㛫ඹᏑᅾẚ⋡ࡢቑຍࡀணࡉࢀࡿࡀࠊ ᐇ㝿ࠊNf2, Sav1, Amotl2 ࡞ࡢ Hippo ⤒㊰ࡢ㑇ఏ Ꮚᑐࡍࡿ gRNA ࡢᏑᅾẚ⋡ࡀ㢧ⴭቑຍࡋ࡚࠾ࡾࠊ ᮏᐇ㦂⣔ࡼࡗ࡚⣽⬊➇ྜ㛵㐃㑇ఏᏊࡢ᳨⣴ࡀྍ⬟ ࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋࡲࡓࠊ㛫ࡶቑຍࡍ - 358 - ࡿ⣽⬊➇ྜࡢ⪅࡞ࡿ᪂つ㑇ఏᏊࡢೃ⿵ࡢࣜࢫࢺ ࠊ㛫ඹῶᑡࡍࡿᩋ⪅࡞ࡿ᪂つ㑇ఏᏊࡢೃ ⿵ࡢࣜࢫࢺࡀᚓࡽࢀࡓࠋࡑࡢ୍᪉࡛ࠊ௨ୗࡢၥ㢟Ⅼ ࡶ᫂ࡽ࡞ࡗࡓࠋ2 ᅇࡢᐇ㦂㛫ࠊ࠶ࡿ࠸ࡣ㸯ࡘࡢ㑇 ఏᏊෆࡢ」ᩘࡢ gRNA 㛫࡛ࠊ⤖ᯝࡢࡤࡽࡘࡁࡀࡁ ࡃࠊቑῶࢆุᐃࡋࡃ࠸㑇ఏᏊࡀከᩘ࠶ࡗࡓࠋࡲࡓࠊ ⣽⬊➇ྜࡢᩋ⪅࡞ࡿ㑇ఏᏊࡢೃ⿵ࡢ୰ࡽ 16 ಶ ࡢ㑇ఏᏊࢆ㑅ࡧࠊಶู㑇ఏᏊᶵ⬟ࢆ☜ㄆࡋࡓࡇ ࢁࠊ10 ಶࡢ㑇ఏᏊ࡛⣽⬊ቑṪࡢపୗࡀぢࡽࢀࠊࡑࡢ ࠺ࡕ 6 ಶࡢ㑇ఏᏊ࡛⣽⬊➇ྜࡢᩋ⪅࡞ࡿࡇࡀ☜ ㄆࡉࢀࡓࠋ6 ಶࡢ㑇ఏᏊࡣᙳ㡪ࡀ࡞ࡃࠊࢫࢡ࣮ࣜࢽࣥ ࢢࡢഇ㝧ᛶ࡛࠶ࡿࡇࡀศࡗࡓࠋࢫࢡ࣮ࣜࢽࣥࢢࡢ ຠ⋡ࢆୖࡆࡿࡓࡵࡣࠊࣛࣥࢲ࣒࡞ࡤࡽࡘࡁࡀࡢ ⛬ᗘ㉳ࡁࡿࡢࢆ᫂ࡽࡋࠊࡑࢀࢆ⪃៖ࡋุ࡚᩿ ࡍࡿᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡓࠋ <2015 ᖺᗘ> 2015 ᖺᗘࡣࠊ๓ᖺࡢၥ㢟ⅬࢆゎỴࡍࡿࡓࡵࠊྛ gRNA ࡢᏑᅾẚ⋡ࡢኚືࡢࣛࣥࢲ࣒࡞ࡤࡽࡘࡁࡢ⛬ ᗘࢆ⪃៖ࡋࡓࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡗࡓࠋࡑࡢࡓࡵࠊ gRNA ࣛࣈ࣮ࣛࣜࡋ࡚ࠊ᪂ࡓࠊ≉ᐃࡢ㑇ఏᏊ ࢆᶆⓗࡋ࡞࠸ non-target gRNA ࢆ 1000 ಶྵࢇ࡛ ࠸ࡿࠊZhang ࡽࡢ gRNA ࣛࣈ࣮ࣛࣜ(Nat Methods 11, 783-784, 2014)ࢆ⏝࠸ࡓࠋࡑࡢ⤖ᯝࠊࡲࡎࠊ4 ᪥ ┠ࡢኚ␗⣽⬊ࣛࣈ࣮ࣛࣜෆࡢᏑᅾẚ⋡ࡀప࠸ gRNA ࡣࣛࣥࢲ࣒࡞ࡤࡽࡘࡁࡢᙳ㡪ࢆࡁࡃཷࡅࡿ ࡇࡀࢃࡗࡓࡓࡵࠊࡑࡢᵝ࡞ gRNA ࡢࢹ࣮ࢱࡣゎ ᯒࡽ㝖ࡋࡓࠋࡉࡽࠊnon-target gRNA ࡢᏑᅾ ẚ⋡ࡣࣛࣥࢲ࣒ኚືࡍࡿࡀࠊ2 ᅇࡢᐇ㦂⤖ᯝࡀඹ ࠶ࡿᩘ್௨ୖቑຍ(࠶ࡿ࠸ࡣῶᑡ)ࡍࡿ☜⋡ࡣప࠸ ࡇࡀศࡗࡓࠋࡑࡇ࡛ࠊࡑࡢ್ࢆᇶ‽ࡋ࡚ࠊ2 ᅇࡢ ᐇ㦂⤖ᯝࡀࠊඹࡑࡢᩘ್௨ୖࡢኚືࢆ♧ࡋ࡚࠸ࡿ ࡶࡢࢆࠊពࡢ࠶ࡿኚືࢆ♧ࡍ gRNA ࡋࡓࠋࡉࡽ ࠊࡑࡢᵝ࡞ពࡢ࠶ࡿኚືࢆ♧ࡍ gRNA ࡀ㸯ࡘࡢ 㑇ఏᏊᑐࡋ࡚」ᩘᏑᅾࡍࡿሙྜࠊࡑࡢ㑇ఏᏊࡀ ⣽⬊➇ྜ㛵ࢃࡿ㑇ఏᏊࡋࡓࠋࡇࢀࡽࡢᇶ‽ࢆ⏝ ࠸ࡿࡇ࡛ࠊ࡞ࡾಙ㢗ᗘࡢ㧗࠸⪅࣭ᩋ⪅ࡢ㑇ఏ Ꮚೃ⿵ࡢࣜࢫࢺࢆᚓࡿࡇࡀ࡛ࡁࡓ⪃࠼࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ࡢᨭࢆཷࡅࡿࡇࡼࡾࠊ࣐࢘ࢫࡢ⣽⬊➇ ྜ㛵㐃㑇ఏᏊࢆ⥙⨶ⓗ᳨⣴࡛ࡁࡿ᪂ࡓ࡞ᐇ㦂⣔ࢆ సࡿࡇࡀ࡛ࡁࡓࠋࡇࢀࡲ࡛ࠊ⣽⬊➇ྜࡢ◊✲ࡣࠊ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ࡢ㑇ఏᏛⓗゎᯒࡀඛ⾜ࡋ࡚࠸ࡿࡀࠊ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚࠾ࡅࡿ᪂つ㑇ఏᏊࡢ᳨⣴ࡣࠊ⣽ ⬊➇ྜ␗ᖖࢆ♧ࡍኚ␗యࢆྠᐃࡋࡓᚋࠊࡑࡢཎ ᅉ㑇ఏᏊࢆྠᐃࡍࡿ࠸࠺᪉ἲࢆ⏝࠸ࡿࡓࡵࠊࡲࡎ ኚ␗యࡢࣜࢫࢺࡀᚓࡽࢀࠊ⣽⬊➇ྜࡢࡼ࠺࡞㑇 ఏᏊࡀ㛵ࢃࡗ࡚࠸ࡿࡢࠊࡑࡢㇺࢆ⌮ゎࡍࡿࡣ 㛫ࡀࡿࠋࡑࢀᑐࡋ࡚ࠊᮏ᪉ἲ࡛ࡣࠊ᭱ึ ࡽ⣽⬊➇ྜ㛵ࢃࡿ㑇ఏᏊࡢࣜࢫࢺࢆᚓࡿࡇࡀ࡛ ࡁࡿࡓࡵࠊᚋࠊᚓࡽࢀࡓ㑇ఏᏊྡࡢࣜࢫࢺࢆࠊ᪤ ▱ࡢࢿࢵࢺ࣮࣡ࢡሗ↷ࡽࡋྜࢃࡏࡿࡇࡼࡾࠊ ⣽⬊➇ྜ㛵ࢃࡿࢩࢢࢼࣝ⤒㊰ࠊ௦ㅰ⤒㊰࡞ࡢ ሗࢆ୍Ẽᚓࡿࡇࡀ࡛ࡁࠊ⣽⬊➇ྜ㛵ࡍࡿ㑇ఏ Ꮚࢿࢵࢺ࣮࣡ࢡࡢᐜࢆಠ▔ࡍࡿࡇࡀ࡛ࡁࡿࡼ࠺ ࡞ࡿᮇᚅࡉࢀࡿࠋࡇࡢࡼ࠺ࠊ⣽⬊➇ྜ㛵ࢃ ࡿ㑇ఏᏊࢿࢵࢺ࣮࣡ࢡࡢయീࢆ⌮ゎࡍࡿࡇࡼ ࡾࠊࡇࢀࡲ࡛㐍ࡵࡽࢀ࡚ࡁࡓࡼ࠺࡞ࠊ≉ᐃࡢಶู㑇 ఏᏊὀ┠ࡋࠊࡑࡢ࿘㎶㡯ࡢゎᯒࢆ㐍ࡵ࡚⾜ࡃ ࠸࠺◊✲ࢫࢱࣝࡣࡃ␗࡞ࡿࠊࢩࢫࢸ࣒ࣞ࣋ࣝ ࡛ࡢ◊✲ࢆ㐍ࡵࡿࡇࡀྍ⬟࡞ࡾࠊ⣽⬊➇ྜࡢ యീࡢ⌮ゎࡀ῝ࡲࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ↓ࡋ - 359 - බເᨭㄢ㢟㸸2013 ᖺᗘ ࢯ࢟ࣀࣜࣥࣝ࢝ࣟࢻ⏕ྜᡂ࣐ࢩࢼ࣮ࣜࡢᵓ⠏ ᪂Ꮫ⾡㡿ᇦ◊✲ బ⸨ ᩥᙪ࣭ி㒔ᏛᏛ㝔⏕⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ᳜≀ࡣከᵝ࡞ḟ௦ㅰ⏘≀ࢆ⏘⏕ࡍࡿࡀࠊࡑࡢ⏕ྜ ᡂ⣔ࡘ࠸࡚ࡣࠊゎ᫂ࡀ㐜ࢀ࡚࠸ࡿࠋ≉ࠊྵ❅⣲ ḟ௦ㅰ⏘≀㸦ࣝ࢝ࣟࢻ㸧ࡣ⏕⌮άᛶࡀ㢧ⴭ࡛ ࠶ࡿࡀࠊࡑࡢ⏘⏕ࡀ≉ᐃࡢ᳜≀✀㝈ᐃࡉࢀࠊࡘࠊ ⤌⧊≉␗ᛶࡀ࠶ࡿࡓࡵࠊ⏕ᏛⓗࠊศᏊ⏕≀Ꮫⓗ ゎᯒࡀᅔ㞴࡛࠶ࡗࡓࠋᡃࠎࡣࠊࢯ࢟ࣀࣜࣥࣝ࢝ ࣟࢻ╔┠ࡋࠊࡑࡢ⏘⏕ᇵ㣴⣽⬊⣔ࢆ☜❧ࡍࡿࡇ ࡼࡾࠊྠ⏕ྜᡂ⣔ࡢゎᯒࢆ㐍ࡵࠊ⏕ྜᡂ㑇ఏᏊ ࡢከࡃࢆ༢㞳ྠᐃࡍࡿࡶࠊ␗✀Ⓨ⌧⣔ࢆ⏝࠸ ࡚ᚤ⏕≀⏕ྜᡂ࣐ࢩࢼ࣮ࣜࢆᵓ⠏ࡍࡿࡇࡶ ᡂຌࡋ࡚࠸ࡓࠋ୍᪉ࠊ⏕ྜᡂ⣔ࡣࠊᮍྠᐃࡢ⏕ྜ ᡂ㑇ఏᏊࡀከᩘᏑᅾࡋࠊࡑࡢ༢㞳ྠᐃࡼࡾࠊࡉࡽ ከᵝ࡞ḟ௦ㅰ≀ࡢ⏕⏘ࡀྍ⬟࡞ࡿᮇᚅࡋࡓࠋ ࡇࢀࡲ࡛ࠊᮍྠᐃࡢ㑇ఏᏊࢆ༢㞳ྠᐃࡍࡿ᪉ἲ ࡋ࡚ࠊ࣓ࢱ࣮࣒࣎ࣟࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࡢ⤫ྜ ⓗゎᯒࡀ᭷ຠ࡛࠶ࡿࡇࢆ᫂ࡽࡋ࡚ࡁࡓࡀࠊࢤ ࣀ࣒㓄ิࢆඖࡋࡓໟᣓⓗ࡞ゎᯒ᪉ἲࡀ࠶ࡿ⪃࠼ ࡓࠋ≉ࠊ㏆ᖺࠊ᳜≀ḟ௦ㅰ⏕ྜᡂ⣔࠾࠸࡚ࡶ 㑇ఏᏊࢡࣛࢫࢱѸࡢᏑᅾࡀ᫂ࡽ࡞ࡗ࡚ࡁࡓࡇ ࡼࡾࠊࢤࣀ࣒㑇ఏᏊ㓄ิࡽࡢ⏕ྜᡂ㓝⣲㑇ఏᏊࡢ ᥈⣴ᶵ⬟ゎᯒࢆ⏝࠸ࡓ᪂ወ࡞⏕ྜᡂ࣐ࢩࢼ࣮ࣜࡢ ᵓ⠏ࢆ᳨ウࡍࡿࡇࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࢣࢩ⛉ࡢࣁࢼࣅࢩࢯ࢘ࡣࠊࡇࢀࡲ࡛ࢯ࢟ࣀࣜࣥ ࣝ࢝ࣟࢻࡢ⏕ྜᡂ㓝⣲㑇ఏᏊࠊ୪ࡧࡑࡢⓎ⌧ ไᚚࡀ⢭ຊⓗゎᯒࡉࢀࡿࡶࠊẚ㍑ⓗࢤࣀ࣒ ࢧࢬࡀᑠࡉ࠸ࡇ㸦⣙ 500Mb㸧ࠊࡲࡓࠊEST ሗ (16537 㓄ิ)ࡀᩚഛࡉࢀ࡚࠸ࡿࡇࡽࠊḟୡ௦ࢩ࣮ ࢣࣥࢧ࣮ࡼࡿࢻࣛࣇࢺࢤࣀ࣒ゎㄞࡀྍ⬟⪃࠼ࡓࠋ ᡃࠎ⮬㌟ࡀணഛⓗ⾜ࡗࡓࢤࣀ࣒ゎᯒࡼࡾ 360 Mb ࡢ㓄ิࡀᚓࡽࢀ࡚࠸ࡓࡀࠊN50 ࡀ 1.7 Kb ▷ ࡗࡓࡇࡼࡾࠊࢤࣀ࣒ᨭࡼࡾࠊࡼࡾ㛗࠸㓄ิ ሗࢆᚓࡿࡇࢆᮇᚅࡋࡓࠋᖹᆒࣥࢧ࣮ࢺࢧࢬ 342bp, 545bp ࡢ PE(࢚࣌ࣥࢻ㸧300, PE500,ᖹᆒ ࣥ ࢧ ࣮ ࢺ ࢧ ࢬ 2820bp, 4,545bp, 7,926bp, 8,945bp, 13,158bp, 15,125bp ࡢ MP(࣓ࢺ࣌)3k, MP5k, MP8k, MP10k, MP15k, MP20k ࣛࣈࣛࣜ ࣮ࢆ࣮ࣜࢻ㛗 150bp ࡎࡘࠊࡑࢀࡒࢀ⣙㸴൨ࠊ⣙㸳൨ࠊ ⣙ 2,2 ൨ࠊ⣙ 2.5 ൨ࠊ⣙ 1.2 ൨ࠊ⣙ 3.6 ൨ࠊ⣙ 1.3 ൨ࠊ ⣙ 1.7 ൨ゎㄞࡍࡿࡶࠊPlatanus ࢯࣇࢺࢆ⏝࠸ ࡓࢵࢭࣥࣈࣝゎᯒࢆ࠾㢪࠸ࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࢤࣀ࣒ゎㄞᨭࡼࡾ 43,738 ࢥࣥࢸࢢ㸦⥲ᘏ㛗ሷ ᇶᩘ 419 Mbࠊ᭱㛗 340 Kbࠊ N50; 21,362 bp㸧ࠊ 15,668 scaffold㸦⥲ᘏ㛗ሷᇶᩘ 467 Mb, ᭱㛗 4.25 Mb, N50; 796 kb㸧ࡢ㓄ิሗࢆᚓࡿࡇࡀ࡛ࡁࡓࠋ ࡇࡢ㓄ิሗࡣࠊࢤࣀ࣒ࢧࢬ 500 Mb ࡍࡿ ࡰ 9 ࢆ࢝ࣂ࣮ࡍࡿࡶࡢ࡛࠶ࡗࡓࠋࡲࡓࠊNCBI Ⓩ㘓ࡉࢀ࡚࠸ࡿ EST ࡢ 9 ࡀ 90%࢝ࣂ࣮⋡࡛࣐ࢵࣉ ࡉࢀࠊࡉࡽࠊPhytometasyn Ⓩ㘓ࡉࢀ࡚࠸ࡿ RNA 㓄ิࡘ࠸࡚ࡶࠊ9 ࢆ㉺࠼ࡿ㓄ิࡀ 50㸣࢝ࣂ࣮⋡ ࡛࣐ࢵࣉࡉࢀ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡾࠊᅇࡢࢤ ࣀ࣒ゎㄞࡼࡾࠊᵓ㐀㑇ఏᏊࡢከࡃࢆ࢝ࣂ࣮ࡍࡿሷ ᇶ㓄ิሗࡀᚓࡽࢀࡓุ᩿ࡉࢀࡓࠋ ࡇࡢࢻࣛࣇࢺ㓄ิሗࢆඖࠊࢯ࢟ࣀࣜࣥࣝ ࢝ࣟࢻ⏕ྜᡂ⣔㑇ఏᏊࢡࣛࢫࢱѸࡢᏑᅾࢆ᳨ウࡋ ࡓࠋṧᛕ࡞ࡀࡽࠊ᫂ⓑ࡞㑇ఏᏊࢡࣛࢫࢱѸࡣㄆࡵࡽࢀ ࡞ࡗࡓࡀࠊ」ᩘࡢࢯ࢟ࣀࣜࣥࣝ࢝ࣟࢻ⏕ྜ ᡂ㛵㐃㑇ఏᏊࡀ㏆᥋ࡋ࡚Ꮡᅾࡍࡿ scaffold ࢆぢࡘࡅ ࡿࡇࡀ࡛ࡁࡓࠋ ࡲࡓࠊࢯ࢟ࣀࣜࣥࣝ࢝ࣟࢻ⏕ྜᡂ⣔࡛㔜せ ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿࢳࢺࢡ࣮࣒ࣟ P450 㑇ఏᏊ㸦௨ ୗࠊP450 ␎ࡍ㸧ࡢ㑇ఏᏊᩘࢆ᥎ᐃࡋࡓࡇࢁࠊ ࢯ࢟ࣀࣜࣥࣝ࢝ࣟࢻ⏕ྜᡂ⣔㛵ࡍࡿ⪃࠼ ࡽࢀࡿ P450ࠊ≉ࠊCYP82 ࣇ࣑࣮ࣜࡀࢻࣛࣇࢺ ࢤࣀ࣒ୖከࡃᏑᅾࡍࡿࡇࠊࡲࡓࠊྠ㑇ఏᏊࣇ ࣑࣮ࣜࡀ≉ᐃࡢ scaffold ෆ࡛㔜」ࡋ࡚Ꮡᅾࡍࡿࡇ ࢆぢฟࡋࡓࠋࡲࡓࠊࡇࢀࡽ CYP82 ࣇ࣑࣮ࣜ㑇ఏᏊ 㛫ࡣࠊⓎ⌧ࡁ࡞㐪࠸ࡀ࠶ࡿࡇࡀุ᫂ࡋࡓࠋ ࡑࡇ࡛ࠊࡇࢀࡽ CYP82 㑇ఏᏊࡢⓎ⌧ࣝ࢝ࣟࢻ ⏘≀ࡢ┦㛵ࢆゎᯒࡋࡓ⤖ᯝࠊࣁࢼࣅࢩࢯ࢘ࡢせ ࣝ࢝ࣟࢻ࡛࠶ࡿࢧࣥࢠࢼࣜࣥࡽࡉࡽỈ㓟 ࡉࢀ࡚⏕ᡂࡍࡿ࣐࢝ࣝࣆࣥࡢ✚┦㛵ࡋࡓⓎ⌧ࢆ ♧ࡍ CYP82 㑇ఏᏊࢆᩘࢡ࣮ࣟࣥぢฟࡋࡓࠋ ࡇࢀࡽࡢ᪂つ P450 㑇ఏᏊࢆ༢㞳ࡋࠊ㓝ẕ࠾࠸ ࡚Ⓨ⌧ࡋࠊࢪࣄࢻࣟࢧࣥࢠࢼࣜࣥࢆᇶ㉁ᛂࢆ⾜ ࡗࡓ⤖ᯝࠊ୍ࡘࡢ᪂つ CYP82 ࡀྠᇶ㉁ࢆỈ㓟ࡍࡿ ࡇࢆㄆࡵࡓࠋࡲࡓࠊูࡢ CYP82 ࡣࠊᆅୖ㒊≉␗ⓗ Ⓨ⌧ࡋࠊࣞࢳࢡࣜࣥࢆᇶ㉁ࡋ࡚ࠊ᪂つ࡞ྜ≀ ࢆ⏘⏕ࡍࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࢀࡽࡢ㑇ఏᏊ ⏘≀ᶵ⬟ࡘ࠸࡚ࡣࠊࡉࡽヲ⣽࡞ゎᯒࡀᚲせ࡛࠶ ࡿࡀࠊࡇࢀࡽ௨እࡶከࡃࡢᮍゎᯒࡢ P450 ࢆぢฟ ࡍࡇࡀ࡛ࡁࠊࣝ࢝ࣟࢻ⏕ྜᡂ⣔ࡢ᪂ࡓ࡞㓝⣲ 㑇ఏᏊࡢⓎ᥀ࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᅇࡢࢤࣀ࣒ゎᯒࡽࠊ᳜≀࠾࠸࡚ࡶࠊࢤࣀ࣒ ሗࢆඖࡋࡓኳ↛≀⏕ྜᡂ⣔ゎᯒࡢ⣒ཱྀࡀぢ࠼ ࡚ࡁࡓࠋ≉ࠊⓎ⌧ࡋ࡞࠸ࡓࡵࠊゎᯒࡀྍ⬟ ࡛࠶ࡗࡓỿ㯲ࡢ⏕ྜᡂ㑇ఏᏊࢆ⏕ྜᡂ࣐ࢩࢼ࣮ࣜ ࡢ᪂ࡓ࡞⣲ᮦࡋ࡚⏝࡛ࡁࡿྍ⬟ᛶࡀ࡛࡚ࡁ࡚ ࠸ࡿࠋࡲࡓࠊⓎ⌧ࡋ࡚࠸ࡿ RNA 㓄ิࡢࢵࢭ ࣥࣈ࣮ࣜࡼࡾࠊⓎ⌧ไᚚᶵᵓࢆ⌮ゎࡍࡿ⣒ཱྀࡀ ࡛࡚ࡁࡓࠋᚑ᮶ࡢࢺࣛࣥࢫࣜࣉࢺ࣮࣒㸭࣓ࢱ࣎ࣟ ࣮࣒ࡢ⤫ྜゎᯒࢆ㉺࠼ࡓ᪂ḟඖࡢኳ↛≀⏕ྜᡂ◊ ✲ࡀྍ⬟࡞ࡿᮇᚅࡋ࡚࠸ࡿࠋࡲࡓࠊḟ௦ㅰ ࡢศᏊ㐍ࡘ࠸࡚ࡶࠊ᭷ຊ࡞▱ぢࡀᚓࡽࢀࠊ ᚋࡢⓎᒎࡀᮇᚅ࡛ࡁࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 బ⸨ᩥᙪ ᳜≀ࡢ⏕㛗ㄪ⠇ 50(2), 93-93 (2015) - 360 - බເᨭㄢ㢟㸸2013 ᖺᗘ ḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ࢆ⏝࠸ࡓᨺᑕ⥺ࡢࣛࢵࢺᮍ⇍༸ẕ⣽⬊ཬࡰࡍ㑇ఏⓗᙳ㡪ホ౯ ᇶ┙◊✲㸦㹁㸧 బ⸨ ᗣᡂ࣭ᨺᑕ⥺ᙳ㡪◊✲ᡤศᏊ⏕≀⛉Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࣄࢺ࠾ࡅࡿᨺᑕ⥺⿕ࡤࡃࡢ⥅ୡ௦ࣜࢫࢡࢆゎ᫂ ࡍࡿࡓࡵࡢື≀ࣔࢹࣝࡋ࡚ࠊ⏨ᛶ⿕ࡤࡃࢆホ౯ࡍ ࡿࡓࡵࡣ࣐࢘ࢫࡀ⏝࠸ࡽࢀ࡚ࡁࡓࠋࡑࢀᑐࡋ࡚ࠊ ዪᛶ⿕ࡤࡃࢆホ౯ࡍࡿື≀ࣔࢹࣝࡋ࡚ࡣࠊ࣐࢘ࢫ ࡣࡑࡢᮍ⇍༸ẕ⣽⬊ࡀᨺᑕ⥺㧗࠸ឤཷᛶ(⣽⬊Ṛ) ࢆ♧ࡋࢇ⣽⬊ࡀᚓࡽࢀ࡞࠸ࡓࡵ㐺ࡉ࡞࠸ࠋࡇ ࡢࡓࡵࠊᙜึࡢィ⏬࡛ࡣࠊᮍ⇍༸ẕ⣽⬊ࡀᨺᑕ⥺ ឤཷᛶࢆ♧ࡉ࡞࠸ࣛࢵࢺࢆື≀ࣔࢹࣝ⏝ࡍࡿண ᐃ࡛࠶ࡗࡓࠋࡋࡋࠊࣛࢵࢺࡢᮍ⇍༸ẕ⣽⬊࠾ࡅ ࡿᨺᑕ⥺ᙳ㡪ࢆ 2 ḟඖ㟁ẼὋືἲࡼࡾㄪࡓ◊✲ ࠾࠸࡚ࠊࣛࢵࢺࡢᮍ⇍༸ẕ⣽⬊࡛ࡢᨺᑕ⥺ㄏⓎ✺ ↛ኚ␗⋡ࡀ࢜ࢫ࣐࢘ࢫࡢ⢭ཎ⣽⬊ẚ࡚࡞ࡾప ࠸ࡇࡀ♧၀ࡉࢀࡓࡓࡵࠊ⥅ୡ௦ࣜࢫࢡࢆゎ᫂ࡍࡿ ࡓࡵḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ࢆ⏝࠸ࡿ᪉ἲࢆᑟධࡍࡿ ࠶ࡓࡗ࡚ࠊ᪤ᨺᑕ⥺ࡼࡾ✺↛ኚ␗ࡀㄏⓎࡉࢀ ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ࣐࢘ࢫᡂ⇍༸ẕ⣽⬊ࢆᶆⓗ⣽ ⬊ࡋࡓࠋࡇࡢሙྜࠊ⢭ཎ⣽⬊↷ᑕࡢሙྜࡢ 2 ಸࡃ ࡽ࠸ࡢ✺↛ኚ␗ࡀ F1 ࣐࢘ࢫほᐹࡉࢀࡿࡇࡀ㐣 ཤሗ࿌ࡉࢀ࡚࠸ࡿࠋ ㏆ᖺࡢḟୡ௦ࢩ࣮ࢣࣥࢫᢏ⾡ࡢ㐍ᒎࡼࡾࠊື≀ ಶయ୍༉ࡈࡢࢤࣀ࣒ࢩ࣮ࢣࣥࢫࡀྍ⬟࡞≧ἣ ࡞ࡗࡓࠋ⥅ୡ௦ࣜࢫࢡࡢゎ᫂ࡢࡓࡵࡣࠊ✺↛ኚ␗ ⋡ࡢపࡉࡢࡓࡵ㠀ᖖከᩘࡢ㑇ఏᏊᗙࢆㄪࡿᚲ せࡀ࠶ࡿࠋࢤࣀ࣒ࢩ࣮ࢣࣥࢫἲ࡛ࡣࠊಶయ୍༉ࡢ ࡰ࡚ࡢ DNA 㓄ิࢆㄪᰝࡍࡿࡇࡀ࡛ࡁࠊሷᇶ⨨ ࠊḞኻ࣭ᤄධࠊࢤࣀ࣒ࡢᵓ㐀␗ᖖ࠸ࡗࡓከࡃࡢ ✀㢮ࡢከᆺࢆゎᯒ࡛ࡁࡿࠋᚑ᮶⏝࠸ࡽࢀࡓᢏ⾡ẚ ࡚ࡣࡿ㧗࠸ゎീᗘ࡛ࡢㄪᰝࢆ⾜࠺ࡇࡀ࡛ࡁ ࡿࡓࡵࠊᚓࡽࢀࡿ⤖ᯝࡣ⏕Ṫ⣽⬊ࡢᨺᑕ⥺↷ᑕ ࡘ࠸࡚ࡢࣜࢫࢡ᥎ᐃ࠾࠸࡚ࢤࣀ࣒య࠾ࡅࡿሷ ᇶᑐ࡛ࣞ࣋ࣝࡢ᭱ࡶヲ⣽࡞ࡶࡢ࡞ࡿணࡉࢀࡓࠋ ᮏ◊✲࡛ࡣࠊ㸲Gy ↷ᑕࡉࢀࡓᡂ⇍༸ẕ⣽⬊⏤᮶ࡢ F1 ࣐࢘ࢫࡢ✺↛ኚ␗ࢆࠊḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ࢆ⏝࠸ ࡓࢤࣀ࣒ࢩ࣮ࢣࣥࢫࡼࡾゎᯒࡋࡓࠋẕぶࡢ↷ ᑕ๓⏘ࡲࢀࡓ F1 ࣐࢘ࢫ↷ᑕᚋ⏘ࡲࢀࡓ F1 ࣐ ࢘ࢫࡢ✺↛ኚ␗ࢆẚ㍑ࡍࡿࡇ࡛ࠊࡑࡢ✀㢮㢖ᗘ ࡘ࠸᳨࡚ウࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᮦᩱ㸸㞤ぶ C57BL/6ࠊ㞝ぶ C3H ࠸࠺⣧⣔ ࣐࢘ࢫࢆ⏝ࡋࡓࠋᨺᑕ⥺་Ꮫ⥲ྜ◊✲ᡤࡢື≀ タ࠾࠸࡚ྠ◊✲ᡤࡢ༠ຊࡢࡶࠊࡲࡎᨺᑕ⥺ࢆ↷ ᑕࡋ࡚࠸࡞࠸ C57BL/6 C3H ࢆ㓄ࡉࡏ F1 ࢆᚓ ࡓࠋึᅇࡢฟ⏘ࡽ㸲㐌ࡀ⤒㐣ࡋࡓᚋࠊ㞤ぶ C57BL/6 4 Gy ࡢ࣐࢞ࣥ⥺ࢆ↷ᑕࡋࡓࠋᡂ⇍༸ẕ ⣽⬊⏤᮶ࡢ F1 ࢆᚓࡿࡓࡵࠊC57BL/6 ࡣ↷ᑕᚋࡍ ࡄ㞝ぶ C3H ྠࡌࢣ࣮ࢪ㸱᪥㛫ධࢀ࡚㓄ࡉ ࡏࡓࠋᨺᑕ⥺↷ᑕ๓ࡢ㓄ࡼࡾ⏘ࡲࢀࡓ F1 ࣐࢘ࢫ 3 ༉ࠊᨺᑕ⥺↷ᑕᚋࡢ㓄ࡼࡾ⏘ࡲࢀࡓ F1 ࣐࢘ ࢫ 3 ༉ࢆᚋࡢゎᯒ⏝࠸ࡓࠋࡑࢀࡒࢀࡢ F1 ࣐࢘ࢫࠊ ぶ࣐࢘ࢫࡢ⭁⮚ࡽᢳฟࡋࡓࢤࣀ࣒ DNA ࢆࢤࣀ ࣒ࢩ࣮ࢣࣥࢫ⏝࠸ࡓࠋ ࢤࣀ࣒ࢩ࣮ࢣࣥࢫ㸸ࡑࢀࡒࢀࡢ࣐࢘ࢫࡢ⭁⮚⏤ ᮶ࢤࣀ࣒ DNA ࢆ⏝࠸࡚ Illumina Hiseq 2000 ࡼ ࡾࢩ࣮ࢣࣥࢫࢆ⾜ࡗࡓࠋ500bp ࡢ DNA ࣇࣛࢢ࣓ࣥ ࢺࣛࣈ࣮ࣛࣜࢆ 100bp ࡢ࢚࣌ࣥࢻἲࡼࡾࢩ࣮ ࢣࣥࢫࢆᐇࡋࡓࠋᚓࡽࢀࡓࢹ࣮ࢱࡣࠊࢤࣀ࣒ᨭ ࡢᨭᢸᙜ⪅ࡢඹྠ◊✲ࡼࡾᅜ❧㑇ఏᏛ◊✲ᡤ ࡢࢫ࣮ࣃ࣮ࢥࣥࣆ࣮ࣗࢱ࣮ࢆ⏝࠸࡚ゎᯒࢆ⾜ࡗࡓࠋ bwa-mem ࢆ⏝࠸࡚࣐࢘ࢫࡢཧ↷㓄ิ(mm10)ࡢ࣐ ࢵࣆࣥࢢࢆ⾜ࡗࡓᚋࠊsamtools GATK ࡼࡾሷᇶ ከᆺࡢᢳฟࢆ⾜ࡗࡓࠋࡑࢀࡒࢀࡢ࣐࢘ࢫ᳨࡛ฟࡉࢀ ࡓሷᇶከᆺࡢ୰࡛ࠊぶࡣ᳨ฟࡉࢀࡎࠊ㸯༉ࡢᏊ౪ ࡢࡳ᳨ฟࡉࢀࡿከᆺࢆ✺↛ኚ␗ࡢೃ⿵ࡋ࡚ࠊࢧ ࣮ࣥ࢞ἲࡼࡿ᳨ドࢆ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ྜィ 8 ༉ࡢ࣐࢘ࢫࡢࢤࣀ࣒ࢩ࣮ࢣࣥࢫゎᯒࢆ⾜ ࡗࡓࠋ୍༉࠶ࡓࡾ⣙ 90 Gb ࡢࢩ࣮ࢣࣥࢫࢆ⾜࠸ࠊF1 ࣐࢘ࢫ㸯༉࠶ࡓࡾ⣙ 500 ಶࡢሷᇶከᆺࡀ᳨ฟࡉࢀ ࡓࠋ ᳨ฟࡉࢀࡓከᆺࡢෆࠊ୧᪉ࡢぶ࣐࢘ࢫࡣ᳨ฟࡉ ࢀࡎࠊ1 ༉ࡢ F1 ࡢࡳ᳨ฟࡉࢀࡿከᆺ࡛ࠊࡢ F1 ࣐࢘ࢫࡣ᳨ฟࡉࢀ࡞࠸ࡶࡢࢆᢳฟࡋࠊࡑࡢ୰࡛ࡶ ࢝ࣂࣞࢵࢪࡀ 15 ௨ୖࠊከᆺࣞࣝࡢ㢖ᗘࡀ 0.35 ࡽ 0.65 ࡞ࡿࡶࡢࢆ de novo ࡢ✺↛ኚ␗ೃ⿵ࡋࡓࠋ 6 ༉ࡢ F1 ࣐࢘ࢫྜィ࡛ 72 ಶࡢሷᇶ⨨ᆺࡢ✺↛ ኚ␗ೃ⿵ࢆᚓࡓࠋ ࡇࡢࡼ࠺࡞✺↛ኚ␗ೃ⿵ࢆࢧ࣮ࣥ࢞ࢩ࣮ࢣࣥࢫ ࡼࡾ☜ㄆࡋࡓࡇࢁࠊඖࡢࣞࣝࡼࡿࣆ࣮ࢡኚ ␗ࢆ♧ࡍࣆ࣮ࢡࡢ㸰ࡘࡢࣆ࣮ࢡࢆ♧ࡍ࣊ࢸࣟ᥋ྜ ࡞ࡗ࡚࠸ࡿᵝᏊࡀほᐹࡉࢀࡓࠋ᳨ドࡢ⤖ᯝࠊሷᇶ⨨ ᆺࡢኚ␗ೃ⿵ࡢෆࠊ47 ಶࡀ┿ࡢ✺↛ኚ␗࡛࠶ࡗࡓࠋ ᨺᑕ⥺↷ᑕ๓ࡢ㓄࡛⏘ࡲࢀࡓ 3 ༉ࡢ F1 ࣐࢘ࢫ 16 ಶࠊ4Gy ࡢ࣐࢞ࣥ⥺ࢆ↷ᑕᚋࡢᡂ⇍༸ẕ⣽⬊⏤᮶ ࡢ F1 ࣐࢘ࢫ㸱༉ 31 ಶࡢሷᇶ⨨ᆺࡢ᪂つ✺↛ኚ ␗ࡀ᳨ฟࡉࢀࠊᨺᑕ⥺↷ᑕࡉࢀࡓ F1 ࣐࢘ࢫ࡛ࡣ✺↛ ኚ␗ࡢᩘࡀቑຍࡋ࡚࠸ࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᨺᑕ⥺ࡼࡾ⏕Ṫ⣽⬊⏕ࡌࡿ✺↛ኚ␗ࡣࠊ࣐࢘ ࢫࡢ≉ᐃ㑇ఏᏊᗙࡘ࠸࡚ࡢ◊✲ࡽḞኻᆺ࡛ ࠶ࡿ⪃࠼ࡽࢀ࡚ࡁࡓࡀࠊࢤࣀ࣒యࡢࡼ࠺࡞ ኚࡀ㉳ࡁ࡚࠸ࡿࡣ࡛᫂࠶ࡗࡓࠋᮏ◊✲ࡼࡾࠊ 4 Gy ࡢ࣐࢞ࣥ⥺ࡀ↷ᑕࡉࢀࡓ࣐࢘ࢫᡂ⇍༸ẕ⣽⬊⏤ ᮶ࡢ F1 ࠾࠸࡚ࠊ↷ᑕ๓ࡢ㓄ࡼࡾ⏕ࡲࢀࡓ F1 ẚ࡚⣙ 2 ಸࡢ᪂つሷᇶ⨨ᆺ✺↛ኚ␗ࡀほᐹࡉ ࢀࡓࠋࡇࡢ⤖ᯝࡣࠊᨺᑕ⥺ࡀᡂ⇍༸ẕ⣽⬊࠾࠸࡚ ሷᇶ⨨ࢆቑຍࡉࡏ࡚࠸ࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 361 - බເᨭㄢ㢟㸸2014 ᖺᗘ 51$ ࢆࡋࡓ᳜≀᪻ࡢ⏕≀⏺ࢆࡲࡓ࠸ࡔ㑇ఏᏊⓎ⌧ไᚚࡢ⌮ゎ ᪂Ꮫ⾡㡿ᇦ◊✲ బ⸨ ㇏࣭ྡྂᒇᏛᏛ㝔⏕㎰Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 㸺ᨭࡢෆᐜ㸼 ࢪࣕ࢞ࣔࡢᐖ࡛࠶ࡿࢽࢪࣗ࢘ࣖ࣍ࢩࢸࣥ ᮏ◊✲࡛ࡣࠊࢪࣕ࢞ࣔࡢ㔜㙐 RNA ࡢ㓄ิ ࢺ࢘ࡢ㣵㔜㙐RNA (dsRNA)ࢆΰࡐ࡚࠾ࡃࠊ ࢽࢪࣗ࢘ࣖ࣍ࢩࢸࣥࢺ࢘ࡢⓎ⌧㑇ఏᏊࡢ㛫࡛┦ ࢚ࢧࡽྲྀࡾ㎸ࡲࢀࡓRNAࡀࢽࢪࣗ࢘ࣖ࣍ࢩࢸ ྠᛶ᳨⣴ࢆ⾜࠸ࠊtrans kingdom interfering RNA ࣥࢺ࢘ࡢ㑇ఏᏊⓎ⌧ࢆไᚚࡋ࠺ࡿࡇࢆࡇࢀࡲ (tkiRNA)ೃ⿵ࢆぢ࠸ࡔࡍࢆ┠ⓗࡋ࡚࠸ࡿࠋ ࡛᫂ࡽࡋࡓࠋ᪻ࡀᦤ㣗ࡼࡾྲྀࡾ㎸ࢇ ᮏᨭ᪂Ꮫ⾡࡛ࡣࠊࢪࣕ࢞ࣔෆᅾࡍࡿ㔜 ࡔdsRNAࡼࡿ᪻ࡢ㑇ఏᏊⓎ⌧ไᚚࡢ⏕≀Ꮫ 㙐 RNA ࢆຠ⋡ⓗᢳฟࡋḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ ⓗ࡞ពࢆ⪃࠼࡚ࡳࡿࠊࢽࢪࣗ࢘ࣖ࣍ࢩࢸࣥ ࡼࡿ RNA-seq ゎᯒࢆᨭ᪂Ꮫ⾡ࠕࢤࣀ࣒ᨭࠖ ࢺ࢘ࡣࢪࣕ࢞ࣔࡀᣢࡘෆᅾᛶdsRNAࢆ࢚ࢧ ࡼࡾ⾜ࡗࡓࠋࡲࡓࠊࢪࣕ࢞ࣔࡢⴥࡶࡋࡃࡣ ࡋ࡚᪥ࠎ㣗࡚࠸ࡿヂ࡛ࠊ᳜≀ࡀᣢࡘෆᅾᛶࡢ ேᕤⓗྜᡂࡋࡓ㔜㙐 RNA ࢆᦤ㣗ࡉࡏࡓࢽ dsRNA㸦ࢇࡣᶵ⬟ᮍ▱㸧ࡀࠊᐇࡣࠊ᪻ ࢪࣗ࢘ࣖ࣍ࢩࢸࣥࢺ࢘ࡢ mRNA-seq ゎᯒࡶ⾜ࡗ ࡞ࡢ㑇ఏᏊⓎ⌧ࢆㄪ⠇ࡋ࡚࠸ࡿࡢ࡛ࡣ࡞࠸ ࡓࠋࡇࡢᐇ㦂ࡼࡾࠊࢪࣕ࢞ࣔࡽࡶࡋ ⪃࠼ࡽࢀࡿࠋᮏ◊✲ࡢ┠ⓗࡣࠊࡇࡢࡼ࠺࡞⏕ tkiRNA ࡀぢࡘࡗࡓሙྜࠊtkiRNA ࡀࡢࡼ ≀⏺ࢆࡲࡓ࠸ࡔdsRNAࡼࡿ㑇ఏᏊⓎ⌧ㄪ⠇ࢆ ࠺࡞ࢱ࣒ࢥ࣮ࢫࡸస⏝ᶵᗎ࡛᪻ࡢ㑇ఏᏊⓎ ⏕≀⏕≀ࡢ┦స⏝ࢆࡾࡶࡘࠕࢤࣀ࣒࣭㑇 ⌧ᙳ㡪ࢆཬࡰࡍࢆ▱ࡿࡀ࡛ࡁࡿࠋ ఏᏊ┦㛵ࠖ⌧㇟ࡢ୍ࡘᤊ࠼ࠊࡑࡢศᏊᶵᵓ࡞ ࡽࡧࡇࡢࡇࡢ⏕≀Ꮫⓗ࡞ព⩏ࢆ᫂ࡽࡍ ࡿࡇ࡛࠶ࡿࠋ RNA ᖸ΅ࡢᘬࡁ㔠ࡶ࡞ࡿ siRNA ࡣከࡃࡢ ┿᰾⏕≀ࡀಖᣢࡍࡿ DICER ࡤࢀࡿ㓝⣲ ࡼࡾ dsRNA ࡽసࡽࢀ࡚࠸ࡿࠋ᳜≀ࡶከᩘࡢ ෆᅾᛶ dsRNA ࡀᏑᅾࡍࡿࡣ▱ࡽࢀ࡚࠸ࡿࡀࠊ ࡑࡢᶵ⬟ࡀࡣࡗࡁࡾ᫂ࡽ࡞ࡗ࡚࠸ࡿࡣ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡇࢀࡲ࡛ࠊࢽࢪࣗ࢘ࣖ࣍ࢩࢸࣥࢺ࢘ࡢ mRNA-seq ゎᯒࢹ࣮ࢱࡘ࠸࡚ࡣࠊࢭࣥࣈࣝ ࢆࡋࠊⓎ⌧ኚື㑇ఏᏊࡢᢳฟࢆ⾜ࡗ࡚࠸ࡿࠋ ᑡ࡞ࡃࡶࠊேᕤྜᡂࡋࡓ㔜㙐 RNA ┦ྠ࡞ 㓄ิࢆᣢࡘࢽࢪࣗ࢘ࣖ࣍ࢩࢸࣥࢺ࢘ࡢෆᅾᛶ㑇 ఏᏊࡢⓎ⌧ࡀࠊᦤ㣗ᚋ 24hr ࡢࢧࣥࣉ࡛ࣝ㢧ⴭ పୗࡍࡿࡇࢆ☜ࡵࡓࠋ ᑡ࡞ࡃࠊከࡃࡀᶵ⬟ᮍ▱࡛࠶ࡿࠋࡇࡢࡼ࠺࡞ dsRNA ࡢᮏ᮶ⓗ࡞ᙺࡀ⏕≀㛫┦స⏝࠶ࡿ ࢆ⏦ㄳ⪅ࡽࡣணࡋ࡚࠾ࡾࠊ⏦ㄳ⪅ࡣᮏ◊✲ ࢆ㏻ࡋ࡚᳜≀ࡀᣢࡘᶵ⬟ᮍ▱ dsRNA ࡢゎᯒ ࡲࡽࡎࠊdsRNA ࢆࡋࡓ⏕≀㛫┦స⏝ ୧⪅ࡢࢤࣀ࣒ඹ㐍㛵ࡍࡿࠊࡃ᪂ࡓ࡞ᴫᛕ ࢆࠕࢤࣀ࣒࣭㑇ఏᏊ┦㛵ࠖࡢ୍ࡘࡋ࡚ᥦࡋࠊ ᮏᴫᛕࡢᇶ♏࣭ᛂ⏝ࡢ୧㠃࠾࠸࡚ࡢᐃ╔ࢆ┠ ᣦࡍࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢪࣕ࢞ࣔࡢⴥࢆᦤ㣗ᚋⓎ⌧ኚືࡍࡿࢽࢪࣗ ࢘ࣖ࣍ࢩࢸࣥࢺ࢘ࡢ㑇ఏᏊࢆᢳฟࡍࡿࡶࠊ ࢪࣕ࢞ࣔࡢ㔜㙐 RNA ࣛࣈ࣮ࣛࣜࡢ㓄ิ ࡽࠊࡇࢀࡽࡢ㑇ఏᏊ┦ྠ࡞㓄ิࡢ᭷↓ࢆ᳨ ⣴ࡍࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ヱᙜ࡞ࡋ - 362 - බເᨭㄢ㢟㸸2014 ᖺᗘ ⏨ᛶዷ࠾ࡅࡿ᪂つཎᅉ㑇ఏᏊࡢྠᐃྥࡅࡓ㸰᪉ྥࡽࡢࣉ࣮ࣟࢳ ᇶ┙◊✲㸦㹁㸧 బ⸨ 㝧୍࣭ᚨᓥᏛᏛ㝔་ṑ⸆Ꮫ◊✲㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ⌧ᅾ᫂ࡽࡉࢀ࡚࠸ࡿ⏨ᛶዷࡢ㑇ఏⓗ⫼ᬒ ࡋ࡚ࠊ< ᰁⰍయୖࡢ↓⢭ᏊᅉᏊ$=)㡿ᇦࡢḞኻ ࡀᣲࡆࡽࢀࡿࠋࡋࡋ↓⢭Ꮚᝈ⪅࡛ $=) 㡿ᇦࡢḞ ኻࡀࡳࡽࢀࡿᝈ⪅ࡣ ࡶ‶ࡓ࡞࠸ࡇࡽࠊ ࡶཎᅉ㑇ఏᏊࡀᏑᅾࡍࡿ⪃࠼ࡽࢀࡿࠋࡑࡇ࡛ࠊ ᡃࠎࡣ᪂ࡓ࡞↓⢭Ꮚཎᅉ㑇ఏᏊࢆྠᐃࡍࡿࡇࢆ ┠ⓗࡋࠊ< ᰁⰍయࢆࢱ࣮ࢤࢵࢺࡋࡓゎᯒࢆ⾜ࡗࡓࠋ < ᰁⰍయࡣ⣙ 㹼0E ࡢࡁࡉ࡛ࠊ୧ᮎ➃ࡣഇᖖ ᰁⰍయ㡿ᇦࡀᏑᅾࡋࠊࡇࡢ㡿ᇦࡣ㑇ఏᏊࡣ↓࠸ ࠸ࢃࢀ࡚࠸ࡿࠋ࣮ࣘࢡ࣐ࣟࢳࣥ㡿ᇦࡣ⣙ 0E ࡽ ࡞ࡾࠊࡇࡢ㡿ᇦෆࡣ ✀㢮ࡢࣃࣜࣥࢻ࣮࣒ࣟ㓄ิ ࡀᏑᅾࡋࠊࡇࡢࣃࣜࣥࢻ࣮࣒ࣟෆ⢭ᕢ࡛Ⓨ⌧ࡋ࡚ ࠸ࡿ㑇ఏᏊࡀከࡃྵࡲࢀ࡚࠸ࡿࡇࡽࠊࡇࡢ㡿ᇦ ⢭Ꮚᙧᡂ㛵ࢃࡗ࡚࠸ࡿ㑇ఏᏊࡀྵࡲࢀ࡚࠸ࡿྍ ⬟ᛶࡀ♧၀ࡉࢀࡿࠋᚑࡗ࡚ࠊࣃࣜࣥࢻ࣮࣒ࣟෆࢆྵ ࡴ < ᰁⰍయ࣮ࣘࢡ࣐ࣟࢳࣥ㡿ᇦࡢኚ␗ࢆ 3DF%LR56 ࡼࡾࣟࣥࢢ࣮ࣜࢻ࡛ゎᯒࡍࡿࡇ࡛ࠊ᪂つࡢ↓⢭ Ꮚཎᅉ㑇ఏᏊࡀྠᐃ࡛ࡁࡿ⪃࠼ࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᑐ㇟ᝈ⪅ࡋ࡚ࠊ㠀㛢ሰᛶ↓⢭Ꮚデ᩿ࡉࢀࠊ 㢧ᚤ㙾ୗ⢭ᕢෆ⢭Ꮚᅇ⾡࠾࠸࡚⢭Ꮚࡀ☜ㄆࡉࢀ ࡎࠊࡑࡢཎᅉ㸦⢭⣴㟼⬦⒗ࠊ᰾ᆺ␗ᖖࠊ$=) 㡿ᇦࡢḞ ኻ࡞㸧ࡀ᫂ࡽࡉࢀ࡚࠸࡞࠸ᝈ⪅ࢆᑐ㇟ࡋࡓࠋ ࢤࣀ࣒ᨭࡼࡾ ࡢ 3DF%LR56 ࡼࡿࢤࣀ࣒ ࢩ࣮ࢣࣥࢫࡀ⾜ࢃࢀࡓࠋ3DF%LR ࡢ࣮ࣜࢻࡣࠊ,OOXPLQD +L6HT ࡼࡾᚓࡽࢀࡓ࣮ࣜࢻ࡛⿵ṇࡉࢀࡓࠋࡲࡓࠊ ,OOXPLQD ࡼࡾᚓࡽࢀࡓ࣮ࣜࢻࢆ UHIHUHQFHJHQRPH KJ࣐ࢵࣆࣥࢢࡋࠊVDPWRROV ࡼࡾ 619ࠊLQGHO ࡀᢳฟࡉࢀࡓࠋ$QQRWDWLRQ ࢆຍᚋࠊGE613EXLOGࠊ 1+/%, ([RPH 6HTXHQFLQJ SURMHFW ࠊ JHQRPHV SURMHFW ࡞ࡢࢹ࣮ࢱ࣮࣋ࢫ࠶ࡿ 619ࠊLQGHO ࡣࣇ ࣝࢱ࣮ࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡢ 3DF%LR56 ࡼࡿࢤࣀ࣒ࢩ࣮ࢣࣥࢫࡼࡾࠊ ᖹᆒ㛗 NE ࡢ࣮ࣜࢻࡀᚓࡽࢀࡓࠋࡋࡋ࡞ࡀࡽࠊ < ᰁⰍయୖࡢࣃࣜࣥࢻ࣮࣒ࣟ㓄ิࡣሷᇶࡢ┦ྠᛶࡶ 㧗ࡃࠊࡲࡓࡑࡢ㛗ࡉࡶ NE㹼0E ࡶཬࡪࡇ ࡽࠊᅇࡢ࣮ࣜࢻࡔࡅ࡛ࡣࠊ㔜」㡿ᇦࢆࣘࢽ࣮ࢡ ⿕そࡍࡿࡇࡀฟ᮶࡞ࡗࡓࠋ୍᪉ࠊ,OOXPLQD ࡼ ࡿࢤࣀ࣒ࢩ࣮ࢣࣥࢫゎᯒࡢ⤖ᯝࠊ᪂つࡢ 619 ࡣ ࠊ LQGHO ࡣ ࡛࠶ࡗࡓ ࠋࡑࡢ࠺ ࡕࠊ QRQV\QRQ\PRXV ࡞᪂つ 619 ࡣ ࠊFRUGLQJ 㡿ᇦࡢ᪂ つ LQGHO ࡣ ࡛࠶ࡾࠊࡇࢀࡽࡢ 619 ࢆྵࡴ㑇ఏᏊࡣ ࠊLQGHO ࢆྵࡴ㑇ఏᏊࡣ ࡛࠶ࡗࡓࠋ< ᰁⰍయ 㛵ࡋ࡚ࡣ 619 ࡀ ࠊ LQGHO ࡀ ࡛࠶ࡗࡓࡀࠊ QRQV\QRQ\PRXV ࡞᪂つ 619 ࡸ FRUGLQJ 㡿ᇦࡢ᪂つ LQGHO ࡣྵࡲࢀ࡚࠸࡞ࡗࡓࠋࡋࡋࠊ᪂つ 619 ࡸ LQGHO ࡢ࠺ࡕࠊ㑇ఏᏊࡢୖὶཬࡧୗὶᏑᅾࡍࡿ࠸ࡃ ࡘࡢኚ␗ࡀࡳࡘࡗࡓࠋࡲࡓࠊ๓㏙ࡢࡼ࠺↓⢭ Ꮚᝈ⪅ࡢ $=) 㡿ᇦࡢḞኻࡀࡳࡽࢀࠊ$=)DE F ࡢ 㡿ᇦࡢ࠺ࡕࠊ≉ $=)D $=)㹠ࡢ㡿ᇦࡢḞኻ ࡣ㢧ᚤ㙾ୗ⢭ᕢෆ⢭Ꮚᅇ⾡ࡼࡿ⢭Ꮚࡢᅇࡀ ྍ⬟࡛࠶ࡿࡇࡽࠊࡇࢀࡽ ࡘࡢ $=) 㡿ᇦෆ↓ ⢭Ꮚࡢཎᅉ㑇ఏᏊࡀᏑᅾࡍࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ ࡑࡇ࡛ࠊࡇࢀࡽ ࡘࡢ $=) 㡿ᇦෆཬࡧ㏆ഐᏑᅾࡍ ࡿ㑇ఏᏊୖࡢኚ␗ࢆࠊ3DF%LR ࡼࡾᚓࡽࢀࡓ࣮ࣜࢻࢆ ⏝࠸࡚ %/$67 ࢧ࣮ࢳࡼࡾ☜ㄆࢆ⾜ࡗࡓࠋ ࡑࡢ⤖ᯝࠊ ᪤▱ࡢ 619 ࡛࠶ࡿࡀࠊ㠀ࢥ࣮ࢻ㡿ᇦࡢኚ␗ࡸ㑇ఏᏊ ࡢୖὶཬࡧୗὶᏑᅾࡍࡿኚ␗ࡀ☜ㄆࡉࢀࡓࠋᚋࠊ ከᩘࡢᖖ⪅↓⢭Ꮚᝈ⪅ࡢ᳨యࢆ⏝࠸࡚ࠊࢧࣥ ࣮࢞ࢩ࣮ࢣࣥࢫࡼࡾࠊࡇࢀࡽࡢ㑇ఏᏊୖࡢኚ␗ࢆ ゎᯒࡋࠊ↓⢭Ꮚࡢཎᅉ㑇ఏᏊࢆྠᐃࡍࡿᚲせࡀ࠶ ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᡃࠎࡣࡇࢀࡲ࡛ࠊ⏨ᛶዷࡢཎᅉ㑇ఏᏊࢆ᫂ࡽ ࡍࡿࡇࢆ┠ⓗࡋࠊ< ᰁⰍయࢆ୰ᚰ◊✲ࢆ⾜ ࡗ࡚ࡁࡓࠋࡑࡢ㛫ࠊ< ᰁⰍయࣁࣉࣟࢢ࣮ࣝࣉࡸ JUJU Ḟኻࡣ↓⢭Ꮚࡸ⢭Ꮚ㐠ື⋡㛵㐃ࡀ࠶ࡿࡇࢆ᫂ ࡽࡋ࡚ࡁࡓࠋࡉࡽ $=)F 㡿ᇦෆᏑᅾࡍࡿ '$=ࠊ %3<ࠊ&'<ࠊ777< ࡞ࡢ㑇ఏᏊࡣࠊ」ᩘࢥࣆ࣮Ꮡᅾ ࡋࠊࡇࢀࡽࡢ㑇ఏᏊࡣ⢭ᕢෆ࡛Ⓨ⌧ࡍࡿࡇࡽࠊ ࡇࢀࡽࡢ㑇ఏᏊࡢࢥࣆ࣮ᩘࡢపୗࡀ⏨ᛶዷࡢཎ ᅉ࡛࠶ࡿ⪃࠼ゎᯒࢆ⾜ࡗ࡚ࡁࡓࠋࡑࡢ⤖ᯝࠊࡇࢀ ࡽࡢ㑇ఏᏊࡢࢥࣆ࣮ᩘ↓⢭Ꮚࡢ㛫ࡣᙉ࠸㛵 㐃ࡀ࠶ࡿࡇࢆ♧ࡋࡓࠋࡋࡋࠊ┿ࡢཎᅉ㑇ఏᏊࢆ ྠᐃࡍࡿࡣ⮳ࡽ࡞ࡗࡓࠋࡲࡓࠊࡇࡢゎᯒࡢ㐣⛬ ࡛㑇ఏᏊࡢḞኻࡀ᩿∦ⓗ࡛࠶ࡿࡇࢆぢฟࡋࡓࠋࡘ ࡲࡾࠊ㛗㙐ࢃࡓࡿࣃࣜࣥࢻ࣮࣒ࣟᵓ㐀ࡢࡓࡵࠊ< ᰁⰍయࡢཧ↷㓄ิࡀ᪥ᮏேࡢ㓄ิࡣྜࡗ࡚࠾ࡽࡎࠊ Ḟኻࢆ᳨ฟࡍࡿ࣐࣮࣮࢝ࡢ⨨ࡀ␗࡞ࡗ࡚࠸ࡿྍ⬟ ᛶࡀ♧၀ࡉࢀࡓࠋᮏᨭࡼࡗ࡚ᚓࡽࢀࡓ 3DF%LR56 ࡼࡿ࣮ࣜࢻ࡛ࡣṧᛕ࡞ࡀࡽ < ᰁⰍయୖࡢࣃࣜࣥࢻ ࣮࣒ࣟ㡿ᇦࢆࣘࢽ࣮ࢡ⿕そࡍࡿࡇࡀฟ᮶࡞ࡗ ࡓࡀࠊᡃࠎࡀゎᯒࢆ⾜ࡗ࡚ࡁࡓ < ᰁⰍయࣁࣉࣟࢢࣝ ࣮ࣉࡸ $=)F 㡿ᇦෆࡢࢥࣆ࣮ᩘ 3DF%LR56 ࡼࡿࣜ ࣮ࢻࢆ⤖ࡧࡘࡅࡿࡇ࡛ࠊ᪥ᮏேࡢ < ᰁⰍయࡢ㓄ิ ࢆ☜ㄆࡍࡿ◊✲ࡢᒎᮃࡀ⪃࠼ࡽࢀࡿࠋ ࡲࡓࠊᮏ◊✲࡛ࡣ < ᰁⰍయࢆࢱ࣮ࢤࢵࢺࡋࡓゎ ᯒࢆ⾜ࡗࡓࡀࠊࢤࣀ࣒ゎᯒࡢᨭࢆᚓࡿࡇࡀฟ ᮶ࡓࡇࡽࠊ< ᰁⰍయ㝈ࡽࡎᖖᰁⰍయࡶᑐ㇟ࡋ ࡚⏨ᛶዷࡢཎᅉ㑇ఏᏊࡢྠᐃ⧅ࡆࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ↓ࡋ - 363 - බເᨭㄢ㢟㸸2010㹼2012 ᖺᗘࠊ &/( ࣌ࣉࢳࢻࢆࣔࢹࣝࡋࡓ᳜≀ࣔࣝࣇ࢛ࢤࣥࡢ㐍స⏝ᶵᵓ㛵ࡍࡿ◊✲ ⱝᡭ◊✲㸦㹑㸧 ⱼ㡬ཬࡧ᰿➃ศ⤌⧊ࡢάᛶไᚚศᏊᶵᵓࡢ┦ྠᛶ┦㐪Ⅼ ᇶ┙◊✲㸦㹀㸧 ⃝ 㐍୍㑻࣭⇃ᮏᏛᏛ㝔⮬↛⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ᙧᡂࡸ⥔ᣢࢆᢸࡗ࡚࠸ࡿࡢࡣࢇศࡗ࡚ ᳜≀ࡣࠊჾᐁࢆษ㝖ࡋ࡚ᇵ㣴ࡍࡿࡇࡸࠊศ ࠸࡞ࡗࡓࠋ ⬟ᛶࢆࡦࡁࡔࡍࡇࡀᐜ࡛᫆࠶ࡿࡇࡽࠊ ᮏ◊✲࡛ࡣࠊࢩࣟࢾࢼࢬࢼࡢ࣌ࣉࢳࢻ࣍ࣝࣔ ࠸ࡶྛჾᐁࠊ⤌⧊ࠊ⣽⬊ࡀ⮬ᚊศᩓⓗ⏕ ࣥࡢ &/9 ࡢస⏝ᶵᵓࡢゎ᫂ࢆ┠ᣦࡋ࡚࠸ࡿࠋᖺ ࡁ࡚࠸ࡿࡼ࠺ぢ࠼ࡿࠋࡋࡋࠊ୰ᯡ⚄⤒⣔ࢆ ᗘࡲ࡛㸱ࡘࡢཷᐜయࡀᶵ⬟ⓗ㛗ᛶࢆࡶࡗ ᣢࡘື≀ࡢࡼ࠺࡞୰ኸ⟶⌮ᆺࡢ⏕ᙧែ࡛ࡣ࡞ ࡚ᶵ⬟ࡍࡿࡇࢆ᫂ࡽࡋ࡚ࡁࡓࡀࠊ+ ᖺ ࠸ࡶࢃࡽࡎࠊྛჾᐁ㸦⣽⬊㸧ࡢ㛫࡛ࡣ☜ ᗘཷࡅࡓࢤࣀ࣒ᨭࡼࡿ✺↛ኚ␗యࡢࣜࢩ࣮ ᐇሗࡢࡸࡾࡾࢆ⾜ࡗ࡚࠸࡚ࠊಶయࡋ࡚ ࢣࣥࢫࡼࡾࠊ ࡑࢀࡒࢀ FOY+ ᖺᗘUSN+ ࡢ⤫୍ᛶࢆಖࡗ࡚࠸ࡿࠋࡑࢀ࡛ࡣࠊࡢࡼ࠺ ᖺᗘFOY+ ᖺᗘࡢ࢚ࣥࣁࣥࢧ࣮✺↛ኚ␗య ࡋ࡚ྛჾᐁ㸦⣽⬊㸧ࡢ㛫࡛ሗࡢࡸࡾࡾࢆ⾜ ࡢཎᅉ㑇ఏᏊࢆྠᐃࡋࡓࠋ ࡗ࡚࠸ࡿࡢࠋࡲࡓࠊྛ⣽⬊ࡣࠊ⣽⬊⮬㌟ࡢ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⨨ሗࡸࠊ⎔ቃࠊⓎ⏕ࣉࣟࢢ࣒ࣛࢆࡢࡼ࠺ FOY ࢚ࣥࣁࣥࢧ࣮✺↛ኚ␗యࡋ࡚ࠊከᩘࡢ✺ ㄆ㆑ࡋ࡚⮬ศ⮬㌟ࢆศࡉࡏಶయࢆᙧసࡗ࡚ ↛ኚ␗యࢆᚓ࡚ࠊᮏࢤࣀ࣒ᨭࡼࡾࠊࢤࣀ࣒ࢩ ࠸ࡿࡢࠋ⌧ᅾࡲ࡛ື≀ࡢ✵㛫ㄆ㆑ࡣᣑᩓ ࣮ࢣࣥࢫࢆ⾜ࡗࡓࠋࡑࡢࢤࣀ࣒ࢩ࣮ࢣࣥࢫ࣐ࢵ ᛶࡢ≀㉁ࡀ㔜せ࡞ᶵ⬟ࢆᣢࡘࡇࡀࡼࡃࡋࡽࢀ ࣆࣥࢢࢆ⤌ࡳྜࢃࡏࡓゎᯒࡼࡾࠊ୕㔞య * ࢱࣥ ࡚࠸ࡿࠋࡋࡋࠊ᳜≀࡛ࡣᙧែᙧᡂ㛵ࢃࡿ࣌ ࣃࢡ㉁ࡢșࢧࣈࣘࢽࢵࢺࠊࣟࢩࣥࣜࢵࢳࣜࣆ࣮ ࢺཷᐜయ࢟ࢼ࣮ࢮ࡛࠶ࡿ %$0&/(1&/(1ཬࡧࠊ ࣉࢳࢻ࣍ࣝࣔࣥࡣࡇࢀࡲ࡛▱ࡽࢀ࡚࠸࡞ࡗࡓࠋ ࣐ࣝࢪࣟࣜࣆ࣮ࢺࢻ࣓ࣥࢆᣢࡕࣘࣅ࢟ࢳࣥࣜ ᡃࠎࡣࠊ&/9 ࡀ ࣑ࣀ㓟ࡢ࣌ࣉࢳࢻ࣍ࣝࣔ ࣮࢞ࢮࡋ࡚ᶵ⬟ࡍࡿ 38% ࢆྠᐃࡋࡓࠋࡉࡽࠊ ࣥࡋ࡚ᶵ⬟ࡍࡿࡇࢆ᫂ࡽࡋ࡚ࡁࡓࡇ ࡑࡢࡢ ✺↛ኚ␗యࡢཎᅉ㑇ఏᏊࡘ࠸࡚ೃ⿵ ࡽࠊᮏ◊✲࡛ࡣࠊࡇࡢ &/9 ࣌ࣉࢳࢻ࣍ࣝࣔࣥ 㑇ఏᏊࡀ≉ᐃ࡛ࡁࡓࠋ ࡢศᏊస⏝ᇶ┙ࡢᵓ⠏ࢆ┠ᣦࡋࡓࠋ ࡇࡢࡼ࠺࡞ࡇࡽࠊᮏᏛ⾡ࢤࣀ࣒ᨭࡼࡾࠊ 㸺ᨭࡢෆᐜ㸼 ከ࡞◊✲ࡢ㐍ᤖࡀ࠶ࡗࡓࠋ㏻ᖖࠊ࣐ࢵࣆࣥࢢ ᡃࠎࡣࠊ᳜≀ࡢ✵㛫ㄆ㆑ᶵᵓゎ᫂ࡢࡓࡵࡢᐇ㦂⣔ ࡽ㑇ఏࡋ༢㞳ࢆ┠ᣦࡍሙྜࠊ㑇ఏᏊᖺࡣࡿ ࡋ࡚᳜≀ࡢⱼ㡬ศ⤌⧊ࢆ୰ᚰࡋࡓศ⤌⧊ ࡇࢁࡀࠊ▷ᮇ㛫㸶ࡘࡢೃ⿵㑇ఏᏊࡀྠᐃ࡛ࡁ ࡢᙧᡂ࣭⥔ᣢᶵᵓὀ┠ࡋ࡚࠸ࡿࠋ⌧ᅾࡢᡤࠊ᳜ ࡓࠋᚋࡶࠊࡉࡽ࡞ࡿゎᯒࡼࡾࠊࢩࢢࢼࣝᅉᏊ ≀ࡢศ⤌⧊ࡢᙧᡂࡸ⥔ᣢࡣࠊ᳜≀࡛࣍ࣝࣔࣥ ࡢྠᐃࡀ㐍ࡴ⪃࠼ࡽࢀࡿࠋ ࠶ࡿ࣮࢜࢟ࢩࣥࢧࢺ࢝ࢽࣥࡀ㔜せ࡞ᶵ⬟ࢆ ᣢࡘࡇࡀ୍⯡ⓗ▱ࡽࢀ࡚࠾ࡾᗈࡃཷࡅධࢀࡽ ࢀ࡚࠸ࡿࠋࡋࡋࠊࡑࢀࡽࡢ࣍ࣝࣔࣥࡀࡢࡼ࠺ ࡞㑇ఏᏊࢆ⏝ࡋࠊࡢࡼ࠺࡞ᶵᵓ࡛ศ⤌⧊ࡢ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ࡢ⤖ᯝࠊࢩࢢࢼࣝఏ㐩ᅉᏊࡀከᩘ༢㞳ࡉࢀ ࡚ࡁࡓࠋᚋࠊࡑࢀࡽࡢᅉᏊࡢ⥘ࡀࡾࢆ᫂ࡽ ࡍࡿࡇ࡛ࠊ᳜≀ࡢᖿ⣽⬊άᛶㄪ⠇ᶵᵓࡢయീ - 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Kinoshita, A., et al., (2015) Plant Signaling & Tabata, R., et al., (2013) Plant Signaling & Behavior. 10:6, e1028707 Behavior. 8. e22534 Kinoshita, A, et al., (2015) Development 142., 444-453. - 365 - බເᨭㄢ㢟㸸2010 ᖺᗘࠊ2012 ᖺᗘࠊ2013 ᖺᗘ ࣐ࣜࣥࣅࣈࣜ࢜ࢆά⏝ࡋࡓᾏ⸴⢾㉁ࡢࣂ࢚࢜ࢱࣀ࣮ࣝኚᢏ⾡ࣉࣛࢵࢺࣇ࢛࣮࣒ࡢᵓ⠏ ᇶ┙◊✲㸦㹀㸧 ࣂ࣐࢜ࢫ⇞ᩱ⏕⏘㐺ࡍࡿ᭱ᑡ㑇ఏᏊࢭࢵࢺࢆᣢࡘ࣐ࣜࣥࣅࣈࣜ࢜ࢭࣝࡢᡂ ᣮᡓⓗⴌⱆ◊✲ ⃝㎶ ᬛ㞝࣭ᾏ㐨ᏛᏛ㝔Ỉ⏘⛉Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 㝣ୖࣂ࣐࢜ࢫࡸῐỈ౫Ꮡࡋ࡞࠸ࣂ࣐࢜ࢫ⇞ᩱ ࡢ⏕⏘ࡋ࡚㸪ᾏὒࣂ࣐࢜ࢫࡢ⇞ᩱኚᢏ⾡ᇶ┙ ࡢຠ⋡ࡀႚ⥭ࡢㄢ㢟࡛࠶ࡿࠋᾏ⸴ࡢ⢾㉁ࢆࣂ࢜ ࢚ࢱࣀ࣮ࣝࡸࣂ࢜Ỉ⣲ኚࡍࡿ≉␗࡞௦ㅰࢆᣢ ࡘ 2 ✀㢮ࡢ࣐ࣜࣥࣅࣈࣜ࢜ࢆぢ࠸ࡔࡋࡓࠋࡇࢀࡽࣅ ࣈࣜ࢜ࡢᾏ⸴⢾㉁ࡽࡢ࢚ࢱࣀ࣮ࣝࡸỈ⣲ࡢ⏕⏘㔞 ࡣ㸪⭠⳦ࡸ㓝ẕࡼࡾࡶ⚽࡛ࡓⓎ㓝⬟ࢆ♧ࡍࠋỈ⏘ ㈨※ࡀ㇏࡞᪥ᮏ࡛ࡣ㸪ᾏ⸴ࡢ⢾㉁ࢆࡣࡌࡵࡍࡿ ᾏὒࣂ࣐࢜ࢫࡣᮍ㛤Ⓨࡢ⏕ྍ⬟࡞࢚ࢿࣝࢠ࣮㈨ ※࡞ࡿࡓࡵ㸪ᮏ◊✲࡛ࡣ㸪ࡇࢀࡽ࣐ࣜࣥࣅࣈࣜ࢜ ࡀᣢࡘ࢚ࢱࣀ࣮ࣝࡸỈ⣲⏕⏘㛵㐃ࡍࡿ௦ㅰ㑇ఏᏊ ⩌ࡢゎᯒ㸦ࢤࣀ࣒ゎᯒ㸧࠾ࡼࡧࡑࡢⓎ⌧ࣂࣛࣥࢫࡢ ゎ᫂ࢆ⾜࠸㸪࣐ࣜࣥࣅࣈࣜ࢜ࡼࡿᾏὒࣂ࣐࢜ࢫ ࡢࣂ࢜⇞ᩱࡢኚᢏ⾡ࡢᇶ┙ࢆᵓ⠏ࡍࡿࠋࡉࡽ 㸪ᾏ⸴≉᭷ࡢ⢾㉁ࢆࣂ࢚࢜ࢱࣀ࣮ࣝࡸࣂ࢜Ỉ ⣲ኚࡍࡿ࣐ࣜࣥࣅࣈࣜ࢜ࢆᑐ㇟ࡋ㸪ࣂ࣐࢜ ࢫ⇞ᩱ⏕⏘㐺ࡍࡿ᭱ᑡࡢ㑇ఏᏊࢭࢵࢺࢆᣢࡘ࣐ࣜ ࣥࣅࣈࣜ࢜ࢭࣝࡢᡂࢆ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࣂ࢜Ỉ⣲⏕⏘ᛶࡢ㧗࠸ Vibrio tritonius AM2 ᰴ ࣜࣇࣞࣥࢫᰴ࡛࠶ࡿ V. harveyi S20 ࡢ࡞ࢤࣀ ࣒ゎᯒࢹ࣮ࢱࢆᚓࡿࡓࡵࡢ fosmid ࣛࣈ࣮ࣛࣜࡢᵓ ⠏ࡑࡢࢩࣙࢵࢺ࢞ࣥࢩ࣮ࢡ࢚ࣥࢫ㸪࠾ࡼࡧࣜࢩ࣮ ࢡ࢚ࣥࢫࡢᨭࢆཷࡅࡓࠋࡲࡓ㸪ࣝࢠࣥ㓟ศゎᛶ ࢞ࢫ⏕⏘ᛶࣅࣈࣜ࢜࡞㸪ィ 11 ✀ࡢࢻࣛࣇࢺࢤࣀ ࣒㓄ิࡢᨭࢆཷࡅࡓࠋࡉࡽ㸪2 ✀㢮ࡢ࣐ࣜࣥࣅ ࣈࣜ࢜㸦V. halioticoli ࠾ࡼࡧ V. tritonius㸧ࢆᑐ㇟ ࡋ࡚㸪␗࡞ࡿ᮲௳ୗ࡛ᇵ㣴ࡋࡓ⳦యࡢ RNA-Seq ሗ ྲྀᚓ࡛ᨭࢆཷࡅࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 I. ࣐ࣜࣥࣅࣈࣜ࢜ࡀᣢࡘࣂ࢜Ỉ⣲⏕⏘⣔㑇ఏᏊᵓ 㐀ࡢࣘࢽ࣮ࢡࡉ V. tritonius AM2 ᰴࡣᾏỈࢆྵࡴᇵ㣴⣔࡛ຠ⋡ࡼࡃ ࣂ࢜Ỉ⣲ࢆ⏕ᡂ࡛ࡁ㸪3 L ࡢᇵ㣴⣔࡛ 10 L ࡢ㉸࠼ ࡿࣂ࢜Ỉ⣲ࡢ⏕⏘ࡀྍ⬟࡞᪂✀ࡢ࣐ࣜࣥࣅࣈࣜ࢜ ࡛࠶ࡿࠋᮏᨭࢆ㏻ࡌ࡚㸪ᮏ⳦ࡢỈ⣲⏕ᡂࡢ㘽㓝⣲ ࡛࠶ࡿࢠ㓟㸫Ỉ⣲࣮ࣜࢮ」ྜయ㸦FHL㸧㑇ఏᏊ⩌ ࡀ 24 kb ࡢ㡿ᇦࢥ࣮ࢻࡉࢀࡿࡇࡀ᫂ࡽ࡞ࡗ ࡓࠋࡉࡽ㸪ᮏᨭ࡛ᚓࡽࢀࡓ 8 ✀ 9 ᰴࡢ࣐ࣜࣥ ࣅࣈࣜ࢜ࡢࢤࣀ࣒ࢹ࣮ࢱࢆẚ㍑ࡋ㸪ձ࢞ࢫ⏘⏕ࣅࣈ ࣜ࢜ࡣ࠸ࡎࢀࡶ⭠⳦࡛ࡣᶵ⬟ࡀᙅ࠸ hyf 㢮ఝࡋ ࡓ㑇ఏᏊᵓ㐀࡛࠶ࡿࡇ㸪ղỈ⣲⏕ᡂ⋡ࡀ㧗࠸ V. tritonius ࡢ FHL 㑇ఏᏊࢡࣛࢫࢱ࣮ࡑࡢࡢάᛶ ࡢప࠸ࣅࣈࣜ࢜ࡢ㑇ఏᏊᵓ㐀ࡀ␗࡞ࡿࡇࡀぢ࠸ࡔ ࡉࢀࡓࠋ II. ᾏ⸴⢾㉁ࡢࣂ࢜⇞ᩱኚᚲせ࡞㑇ఏᏊ⩌ࡢ Ⓨ⌧ືែࡢᢕᥱ ୧⳦ᰴ࠾࠸࡚㸪ᾏ⸴⢾㉁ࢆ௦ㅰࡋ࡚࠸ࡿ⣽⬊ࡢࢤ ࣀ࣒࣡ࢻ࡞㑇ఏᏊⓎ⌧ࡢ୍➃ࡀ᫂ࡽࡍࡿࡇ ࡀ࡛ࡁࡿࡼ࠺࡞ࡗࡓࠋࡲࡎ㸪V. halioticoli ࡛ࡣ㸪 ࢤࣀ࣒ゎᯒࡢࡳ࡛ࡣྠᐃࡀ࡛ࡁ࡞ࡗࡓ㸪ࣝࢠࣥ 㓟ศゎ≀ࡢྲྀࡾ㎸ࡳ㛵ࡍࡿ㑇ఏᏊࢆ⤠ࡿࡇࡀ࡛ ࡁࡓࠋࡇࢀࡽࡢᡂᯝࢆᇶ㸪ᾏỈᏑᅾୗ࡛ࣝࢠࣥ 㓟 ࡽ ࢚ ࢱ ࣀ ࣮ ࣝ ࢆ ⏕ ᡂ ྍ ⬟ ࡞ ௦ ㅰ ᨵ ኚ V. halioticoli ᰴࡢసฟᡂຌࡋࡓࠋ⌧Ⅼ࡛㸪ࡇࢀࡣ ࣝࢠࣥ㓟ࡽ࢚ࢱࣀ࣮ࣝࢆ⏕ᡂྍ⬟࡞᭱ᑠ㑇ఏᏊࢭ ࢵ ࢺ ࢆ ᭷ ࡍ ࡿ ࣐ ࣜ ࣥ ࣅ ࣈ ࣜ ࢜ ࡛ ࠶ ࡿ ࠋ ୍ ᪉ 㸪 V. tritonius ࡛ࡣ㸪24 kb ࠾ࡼࡪࡇࡢ FHL 㑇ఏᏊࢡࣛ ࢫࢱ࣮ࡢ㌿༢ࡀྍど࡛ࡁࡿࡼ࠺࡞ࡾ㸪༢୍ ࡢ㌿⏘≀࡛ࡣ࡞ࡃ㸪3㹼4 㡿ᇦศࢀ࡚Ⓨ⌧ࡉࢀ ࡚࠸ࡿࡶࡢ♧၀ࡉࢀࡓࠋࡉࡽ㸪ᮏ⳦ࡣࢽࢺࣟࢤ ࢼ࣮ࢮ㑇ఏᏊࢡࣛࢫࢱ࣮ࢆ᭷ࡋ㸪ࡇࢀࡀỈ⣲⏕ᡂ ᐤࡋ࡚࠸ࡿ᫂ࡢࡲࡲ࡛࠶ࡗࡓࡀ㸪ᮏᨭࡼ ࡾ㸪㑇ఏᏊࡢⓎ⌧ࡣࢇほᐹࡉࢀࡎ㸪ࡑࡢᐤ ࡣᴟࡵ࡚ప࠸ࡇࡀ♧၀ࡉࢀࡓࠋ III. ࣅࣈࣜ࢜ࡢ㐍⣔⤫ᶞࡢ᭦᪂ᚲせ࡞ሷᇶ㓄ิ ሗࡢྲྀᚓ ࣅࣈࣜ࢜⛉ 78 ✀ࡢᶆ‽ᰴࢆ 9 ࡘࡢࣁ࢘ࢫ࣮࢟ࣆࣥࢢ 㑇ఏᏊᇶ࡙ࡁศᏊ⣔⤫ゎᯒࢆ⾜࠸㸪ࡇࢀࡽࡣ 14 ✀ࡢࢡ࣮ࣞࢻศࡅࡽࢀࡿࡇࢆ♧ࡋࡓࠋࡋࡋ㸪 ᮍࡔከࡃࡢࣅࣈ࡛ࣜ࢜㸪㑇ఏᏊኚ␗ࡢࡁࡉࡽゎ ᯒᑐ㇟ࡍࡿࣁ࢘ࢫ࣮࢟ࣆࣥࢢ㑇ఏᏊࡢቑᖜࡀᅔ㞴 ࡛࠶ࡗࡓࠋᮏᨭࡼࡾ㸪ࡇࡢ࠺ࡕࡢ୍㒊ࡢ✀㸦 V. aerogenes, V. gazogenes, V. rhizosphaerae ࡞㸧ࡢ ሷᇶ㓄ิྲྀᚓᡂຌࡋ㸪ࣅࣈࣜ࢜ࡢ㐍⣔⤫ᶞࢆ ᭦᪂ࡋ㸪᪂ࡓ࡞ࢡ࣮ࣞࢻࢆᥦࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 H21 ࡽ 3 ᖺᗘࢃࡓࡾࢤࣀ࣒ᨭࢆཷࡅࡓ⤖ᯝ㸪 ձಶูࡢ㑇ఏᏊゎᯒ࡛ࡣぢฟࡍࡇࡀᅔ㞴࡛࠶ࡗࡓ ࣐ࣜࣥࣅࣈࣜ࢜ࡢⓎ㓝௦ㅰ㑇ఏᏊࢆ᥈⣴ࡋ㸪ࣅࣈࣜ ࢜⣽⬊ࡢࣂ࢜⇞ᩱ⏕ᡂ㛵ࢃࡿ௦ㅰࢿࢵࢺ࣮࣡ࢡ ࢆ᥎ᐃࡍࡿࡇࡀྍ⬟࡞ࡾ㸪ղࡇࡢ௦ㅰࢿࢵࢺ࣡ ࣮ࢡࢆᇶᾏ⸴⢾㉁ࡢ௦ㅰ㐣⛬࠾ࡅࡿ㑇ఏᏊࡢⓎ ⌧ືែࢆㄪࡿࡇࡀ࡛ࡁࡿࡼ࠺࡞ࡾ㸪ճ࣐ࣜࣥ ࣅࣈࣜ࢜⣽⬊ࢆ⏝࠸࡚㸪ࣂ࢜⇞ᩱ⏕ᡂᴟࡵ࡚ ࡛࠶ࡗࡓࠕࣝࢠࣥ㓟ࠖ࠸࠺ከ⢾ࡽ࢚ࢱࣀ࣮ ࣝࢆ⏕ᡂྍ⬟࡞௦ㅰᨵኚᰴࢆస〇ࡍࡿࡇᡂຌࡋ㸪 ᭷⏝≀㉁ࡢ⏕⏘ᕤሙࡍࡿࡓࡵࡢከᙬ࡞◊✲㸦Vibrio - 366 - Factory ◊✲㸧ࡢᒎ㛤⤖ࡧࡘࡅࡿࡇࡀ⌧ᐇࡢ ࡶࡢ࡞ࡗ࡚ࡁࡓࠋࡉࡽ㸪⏦ㄳ⪅ࡀᪧ᮶ࡽ㐍ࡵ ࡚࠸ࡿࣅࣈࣜ࢜ࡢ⣔⤫㐍ࡢᐜゎ᫂ྥࡅ࡚㸪᪂ ࡓ࡞▱ぢࢆ㞟✚ࡍࡿࡇࡀ࡛ࡁ㸪update ࡋࡓሗࢆ Ꮫ⾡ࢥ࣑ࣗࢽࢸ࣮Ⓨಙࡍࡿࡇࡶ࡛ࡁࡓࠋࣅࣈ ࣜ࢜ࡢࢤࣀ࣒ሗ㛵ࡋ࡚ࡣ㸪140 ✀࡚ࡢሗࢆ ຍࡋࡓ⣔⤫ࡸ㐍ࡢࢢ࣮ࣝࣉศࡅࢆど㔝ධࢀ㸪 ࢤࣀ࣒ሗࡢྲྀᚓ㗦ពດຊࢆࡋ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Sawabe, T. et al., Front. Microbiol. 4, 414 (2013) Matsumura, Y. et al., Int. J. Hydrogen Energy 40, 9137-9146 (2015). - 367 - බເᨭㄢ㢟㸸2010 ᖺᗘ ⏕≀་Ꮫ◊✲᥎㐍ࡢࡓࡵࡢ࢝ࢽࢡࢨࣝ㹋㹆㹁ከᆺሗᇶ┙ࡢᩚഛ ᇶ┙◊✲㸦㹀㸧 ᳝ྡ 㝯࣭ᮾᾏᏛ་Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࢝ࢽࢡࢨࣝ㸦Macaca fascicularis㸧ࡣ iPS ⣽⬊ ࡸ⮚ჾ⛣᳜◊✲ࡢࣔࢹࣝື≀ࡋ࡚⏝ࡉࢀ࡚࠸ࡿ ࡀࠊࢻࢼ࣮ࣞࢩࣆ࢚ࣥࢺࡢ MHC ᆺࢆ⪃៖ࡋࡓ◊ ✲ሗ࿌ࡣᅜෆ࡞࠸ࠋࡼࡗ࡚࢝ࢽࢡࢨࣝࢆ⏝࠸ࡓ 㠀⮫ᗋヨ㦂ࢆ㐍ࡵࡿࡓࡵࡣࠊࡑࡢ MHC ከ ᆺࢆ⌮ゎࡋࠊ≉ᐃ MHC ᆺࡢ࣍ࣔ᥋ྜయࡑࡢ࣊ࢸ ࣟ᥋ྜయࡢᏳᐃ౪⤥యไࢆ☜❧ࡍࡿࡇࡀᚲせ࡛࠶ ࡿࠋࡇࢁࡀ࢝ࢽࢡࢨࣝ MHC㸦Mafa㸧㡿ᇦࡣࠊ 㑇ఏᏊ㔜」ࡼࡾᙧᡂࡉࢀࡓ㢮ఝᛶࡢ㧗࠸ MHC 㑇 ఏᏊࡀᩘከࡃ⨨ࡋ࡚࠾ࡾࠊࣄࢺྠᵝ࡞ከᆺゎᯒ ἲ࡛ࡣࡑࡢከᆺᛶࡢ᫂☜ࡣᅔ㞴࡛࠶ࡿࠋࡑࡇ࡛ࠊ ᮏ◊✲ㄢ㢟࡛ࡣࠊRoche GS FLX ࢩ࣮ࢡ࢚ࣥࢧ࣮ࢆ ⏝࠸ࡓ Mafa ࢡࣛࢫϨ㑇ఏᏊࡢࢩ࣮ࢡ࢚ࣥࢩࣥࢢ ࡘ࠸࡚ࡢᨭࢆཷࡅࠊࡑࡢᚋࡢᒎ㛤ࢆྵࡵ࡚ Mafa 㡿ᇦ࠾ࡅࡿ MHC ࣁࣉࣟࢱࣉᵓ㐀ࢆ᫂ࡽ ࡍࡿࡇࢆ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࢝ࢽࢡࢨࣝ MHC (Mafa) ࢡࣛࢫ I 㑇ఏᏊ⏤᮶ RT-PCR ⏘≀ (400 bp㹼500 bp)ࠊ300 ࢧࣥࣉࣝࡢ Roche GS FLX ࢩ࣮ࢡ࢚ࣥࢧ࣮ࡼࡿࣃࣟࢩ࣮ࢡ࢚ࣥࢩ ࣥࢢ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㸯㸬Mafa 㑇ఏᏊࡢከᆺゎᯒࣁࣉࣟࢱࣉゎᯒ ࢩ࣮ࢡ࢚ࣥࢩࣥࢢࡢᨭࡼࡾࠊィ 112 ✀㢮ࡢ Mafa ࢡࣛࢫ I 㑇ఏᏊࡢሷᇶ㓄ิࢆ᳨ฟࡋࡓࠋࡑࢀࡽ ࡢෆࠊ58 ✀㢮ࡣ᪂つྠᐃࡉࢀࡓࡶࡢ࡛࠶ࡗࡓࠋࡇ ࢀࡽሷᇶ㓄ิࡢ⤌ࡳྜࢃࡏࡽ 17 ✀㢮ࠊ23 ✀㢮ࠊ 12 ✀㢮ࡢ Mafa-AࠊMafa-B/I ࠾ࡼࡧ Mafa-E ࣁࣉࣟ ࢱ ࣉ㸦 Hp 㸧ࢆ ࡑࢀࡒࢀ ᥎ᐃࡋ ࡓࠋ ࡾࢃ ࡅࠊ F/A/E/B-Hp1 ࡸ F/A/E/B-Hp1 ࡢ 㢖ᗘࡣࡑࢀࡒࢀ 10.5%, 10.2%࡛࠶ࡾࠊࡢࣁࣉࣟࢱࣉ㢖ᗘࡼࡾࡶ 㧗࠸್ࢆ♧ࡋࡓࡇࡽࠊ≉ᐃࡢ Mafa ᆺࢆ᭷ࡍࡿ ࣍ࣔ᥋ྜయ࡞ࡢಶయࡣࠊࡇࢀࡽ㧗㢖ᗘࣁࣉࣟࢱ ࣉ↔Ⅼࢆ⤠ࡿࡇࡼࡾຠᯝⓗ᳨ฟࡉࢀࡿ⪃ ࠼ࡽࢀࡓࠋ 㸰㸬NGS ࡼࡿ Mafa 㑇ఏᏊࡢࢪ࢙ࣀࢱࣆࣥࢢἲࡢ 㛤Ⓨ ๓㡯ࡢከᆺሗࢆ⏝࠸࡚ࠊḟୡ௦ࢩ࣮ࢡ࢚ࣥࢩࣥ ࢢἲᇶ࡙ࡃṇ☜ࠊ⡆⣲ࠊ㎿㏿࡞ࢪ࢙ࣀࢱࣆࣥࢢ ἲࢆ㛤Ⓨࡋࡓࠋࡍ࡞ࢃࡕࠊ Mafa-A, -B, -I, -E, -F, -DRB, -DQA1, -DQB1, -DPA1, -DPB1 ࡢከᆺゎᯒ ࡽᚓࡽࢀࡓィ 188 ✀㢮ࡢሷᇶ㓄ิࡘ࠸࡚ࠊࡑࢀࡽ ಖᏑᛶࡢ㧗࠸㒊࣮ࣟ࢝ࢫ≉␗ⓗࣉ࣐࣮ࣛࢆタ ィࡋࠊ࣐ࣝࢳࣉࣞࢵࢡࢫ PCR ἲࡢ᮲௳᳨ウࢆ⾜ࡗࡓࠋ ᮏἲࡢ᳨ドࡋ࡚ࠊ๓㡯⏝࠸ࡓ 127 㢌ࡘ࠸࡚࣐ ࣝࢳࣉࣞࢵࢡࢫ PCR ࢆ⾜࠸ࠊGS Junior ࡸ Ion PGM ࢆ⏝࠸ࡓࣥࣉࣜࢥࣥࢩ࣮ࢡ࢚ࣥࢩࣥࢢࡼࡾ࣮ࣜ ࢻ㓄ิࢆᚓ࡚ࠊ᪤▱ࣜࣝ㓄ิࢆཧ↷ࡋ࡞ࡀࡽࣜ ุࣝᐃࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊ᪤▱ሗ▩┪࡞ࡃุ ᐃࡉࢀࡓࡇࡽࠊᮏἲࡣࠊከ᳨యࡢࢪ࢙ࣀࢱࣆ ࣥࢢࡢࡓࡵࡢඃࢀࡓࢶ࣮࡛ࣝ࠶ࡿ⪃࠼ࡽࢀࡓࠋ 㸱㸬MHC ሗຍ࢝ࢽࢡࢨࣝࡢ⏕་⒪࣭⛣᳜◊ ✲ࡢᛂ⏝ ๓㡯࡚㛤Ⓨࡋࡓࢪ࢙ࣀࢱࣆࣥࢢ᪉ἲࢆ⏝࠸࡚ 5,000 㢌つᶍࡢࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡗࡓ⤖ᯝࠊ47 㢌 ࡢ MHC ࣍ࣔ᥋ྜయࢆࡇࢀࡲ᳨࡛ฟࡋࡓࠋࡑࢀࡽ ࡢෆࡢ 5 㢌ࡣࠊA-Hp8.1ࠊE-Hp2ࠊB/I-Hp1ࠊ#7 ࢆ ᭷ࡍࡿ࣍ࣔ᥋ྜయ࡛࠶ࡗࡓࠋ」ᩘ㢌ࡢ࣍ࣔ᥋ྜయ ࡽࡢ iPS ⣽⬊ࡀ᪤ᶞ❧ࡉࢀ࡚࠾ࡾࠊࢻ࣮ࣃ࣑ࣥศ Ἢ⚄⤒⣽⬊ࠊᚰ➽⣽⬊ࠊ⥙⭷Ⰽ⣲ୖ⓶⣽⬊࡞ࡢ ศᐇ㦂࡞ࡽࡧ⛣᳜ᐇ㦂⏝ࡉࢀ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 iPS ⣽⬊⏤᮶ศ⣽⬊ࡢ⛣᳜ࡸࠊᏊᐑ➼ࡢ⮚ჾ⛣ ᳜ᐇ㦂࠾࠸࡚ࠊᮏ◊✲ㄢ㢟࡚㛤Ⓨࡋࡓ MHC ⤫ ᚚ࢝ࢽࢡࢨࣝࡀ⏝࠸ࡽࢀࡘࡘ࠶ࡿࠋᚋࡶከタ ࡽࡢ㟂せࡀぢ㎸ࡲࢀ࡚࠸ࡿࡇࡽࠊⓎ⏕ᕤᏛⓗ ᡭἲࢆ㥑ࡋࡓேᕤ⦾Ṫἲࡸࢧࣝࡢ⫱ᡂタ࠾ࡅ ࡿ DNA ࢱࣆࣥࢢࢆࡋࡓ࢝ࢽࢡࢨࣝࢆぶࡋ ࡓ⦾Ṫࢥࣟࢽ࣮ࡢస〇ࢆ㐍ࡵࡿ࡞ࠊᏳᐃ౪⤥యไ ࡢᵓ⠏ࢆ㐍ࡵ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ㄽᩥࣜࢫࢺ㸼 Shiina T. et al., Immunogenetics 63, 485-499 (2011) Arikata M. et al., PLoS One 7, e37220 (2012) Blancher A. et al., Immunogenetics 64, 605-614 (2012) de Groot NG. et al., Immunogenetics 64, 615-631 (2012) Shiina T. et al., Tissue Antigens 80, 305-316 (2012) Morizane A. et al., Stem Cell Reports 1, 1-10 (2013) Blancher A. et al., Immunogenetics 66, 219-230 (2014) Kisu I. et al., J Obst Gynaecol Res. 40, 907-918 (2014) Kono A. et al., J Immunol. 192, 3239-3246 (2014) Kisu I. et al., Int J Pharmacol. 40, 907-918 (2014) Shiina T. et al., Immunogenetics 67, 563-578 (2015) Morita D. et al., Nature Communications In press. Kawamura T. et al., Stem Cell Reports In press. - 368 - බເᨭㄢ㢟㸸2012 ~ 2013 ᖺᗘ ࢝ࢥࡢ⬇ఇ╀ㄏᑟ࠾ࡅࡿ⎔ቃ ᗘሗࡢಖᏑ࣭グ᠈ࡢศᏊゎᯒ ᇶ┙◊✲㸦㹀㸧 ሷぢ 㑥༤࣭ಙᕞᏛ⧄⥔Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࢝ࢥ%RPE\[PRULࡢ⬇ఇ╀ㄏᑟࡳࡽࢀࡿᏘ ⠇㐺ᛂࡣぶᏊ㸰ୡ௦ࢃࡓࡗࡓ⊂⮬ࡢ⎔ቃᛂ⟅ᶵᵓ ࡛࠶ࡿࠋ⬇ࢆΥಖㆤࡍࢀࡤḟୡ௦༸ࡣఇ╀ࡋࠊ ⬇Ⓨ⏕ึᮇ࡛ᙧែᙧᡂࡀṆࡍࡿࠋΥ࡛ࡣḟୡ௦ ࡣ㠀ఇ╀༸࡞ࡾࠊ⣙㸯㐌㛫࡛ᗂࡀᏤࡍࡿࠋࡇ ࡢࡼ࠺ࠊぶࡀཷࡅࡓ ᗘ᮲௳ࡼࡾࠊᏊ౪㸦⬇㸧 ࡢ㐠㸦⾲⌧ᆺ㸧ࡀࣜࣉࣟࢢ࣑ࣛࣥࢢࡉࢀࡿࠋࡇࢀ ࡲ࡛ࡢ◊✲࡛ࠊ7537UDQVLHQWUHFHSWRUSRWHQWLDO ࢳࣕࢿࣝࡢ%P753$ࡀ⬇ᮇ࠾࠸࡚Υ௨ୖࡢ ᗘࢆཷᐜࡋࠊḟୡ௦࡛ࡢఇ╀ࢆㄏ㉳ࡍࡿࢭࣥࢧ࣮ศ Ꮚ࡛࠶ࡿࡇࢆ✺ࡁṆࡵࡓࠋࡲࡓࠊఇ╀࣍ࣝࣔࣥ '+ࡣ⻈ᮇࡢ༸ᕢࡢ'+ཷᐜయస⏝ࡋ࡚ఇ╀༸ ࢆㄏᑟࡍࡿ⚄⤒࣌ࣉࢳࢻ࡛࣍ࣝࣔࣥ࠶ࡿࡀࠊ%P753$ ࡢάᛶࡣ'+ࡢ⾑ᾮ୰ࡢ✚ᴟⓗ࡞ᨺฟࢆಁ㐍ࡍ ࡿࡇࡶ᫂ࡽࡋࡓࠋ࠾ࡑࡽࡃࠊ࢝ࢥࡢఇ╀ㄏ ᑟࡣΥ௨ୖࡢ⎔ቃ ᗘࢆཷᐜࡋάᛶࡋࡓ %P753$ࡼࡾࠊ'+ᨺฟࡢไᚚ⧅ࡀࡿ⣙㸯ࣨ᭶ ࠾ࡼࡪ⎔ቃሗࡢಖᏑ࣭グ᠈ࢆక࠺ศᏊࢿࢵࢺ࣮࣡ ࢡྍረⓗኚࡀࡶࡓࡽࡉࢀࠊఇ╀ᛶࢆỴᐃࡋ࡚࠸ ࡿ⪃࠼ࡽࢀࡿࠋࡉࡽࠊ⬇ᮇࢆΥಖㆤࡋࡓሙ ྜࡣග㸦᪥㛗㸧᮲௳ࡼࡾఇ╀ᛶࡀỴᐃࡉࢀࡿࡇ ࡸᗂᮇࡢᰤ㣴᮲௳ࡼࡗ࡚ఇ╀ᛶࡀኚ᭦ࡉࢀࡿ ࡇࡶ▱ࡽࢀ࡚࠸ࡿࠋᮏ◊✲࡛ࡣࠊ࢝ࢥࡢ⬇ఇ╀ ㄏᑟぢࡽࢀࡿⓎ⏕ྍረᛶ࠾ࡅࡿ⎔ቃሗࡢಖ Ꮡ࣭グ᠈ࡢศᏊᐇయࡢゎ᫂ࢆ┠ᣦࡋ࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᮏᨭ࡛ࡣࠊࡇࡢᴫᛕⓗ௬ᐃࡋࡓሗࡢಖᏑ࣭ グ᠈ࡢศᏊᐇయࢆゎ᫂ࡍࡿࡓࡵࠊ᪂ᆺࢩ࣮ࢣࣥࢧ ࡼࡿࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒51$VHTゎᯒࢆ⾜ ࡞࠸ࠊఇ╀ㄏᑟ㛵㐃㑇ఏᏊࡢ⥙⨶ⓗ࡞ࢫࢡ࣮ࣜࢽࣥ ࢢᶵ⬟ゎᯒࢆ㐍ࡵࡓࠋゎᯒ࡛ࡣࠊ㕥ᮌ✨ඛ⏕ࢆࡣ ࡌࡵࡍࡿᮾⳢ㔝◊✲ᐊࢩ࣮ࢣࣥࢫࢳ࣮࣒ࡼࡾ ࢩ࣮ࢡ࢚ࣥࢩࣥࢢࢆࠊᮾிᕤᴗᏛࡢ㯮ᕝ㢧◊✲ᐊ ࡼࡾࢹ࣮ࢱゎᯒࢆ⾜࡞ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᖺᗘࡢᨭ࡛ࡣࠊྜィ᳨యࡢゎᯒࢆ⾜࡞ ࡗࡓࠋ᳨యࡢෆヂࡣࠊࡲࡎΥ࣭ᬯࠊ Υ࣭ᬯࠊ࠾ࡼࡧΥ࣭ᬯ࡛753$ࡢ ࣀࢵࢡࢲ࢘ࣥฎ⌮㸦㸰✀㢮㸧ࢆ⾜࡞࠸ࠊḟୡ௦ࡢఇ ╀ᛶࢆ㐠࡙ࡅࡓ⬇ࡽ51$ࢆᢳฟࡋࠊ51$VHT ゎᯒࢆ⾜࡞ࡗࡓࠋࡲࡓࠊ⬇ᮇࢆΥ࣭ᬯࠊ Υ࣭ᬯಖㆤࡋࡓ⻈ࡢ⬻̾㣗㐨ୗ⚄⤒⠇」ྜయ ࡽᢳฟࡋࡓ51$ࢆ⏝࠸࡚ࠊ51$VHTゎᯒࢆ⾜࡞ࡗ ࡓࠋࡉࡽᖺᗘࡢᨭ࡛ࡣග㸦᪥㛗㸧ࡸᰤ㣴 ᮲௳ࡢ㐪࠸ࡼࡿ51$VHTゎᯒࢆ⬇Ꮚᗂࡸ⻈ ࡢ⬻6*࡛⾜࡞࠸ࠊࡑࢀࡒࢀࡢ⎔ቃ᮲௳࡛Ⓨ⌧㔞ࡢ ␗࡞ࡿ㑇ఏᏊࡢྠᐃࢆヨࡳࡓࠋࡉࡽࠊᖺᗘࡢ ⤖ᯝ↷ࡽࡋྜࢃࡏࡿࡇࡼࡾࠊఇ╀ᛶࡢ㐪࠸ ࡼࡾඹ㏻ࡢᣲືࢆ♧ࡍ㑇ఏᏊࡢྠᐃࢆヨࡳࡓࠋ ᖺᗘࡢᨭࡼࡾࠊࠥࡢ⬇࠾࠸࡚ ࡣࠊ࡛ಶࠊ࡛ಶࠊ ࡛ಶࠊ࡛ಶࠊ ࡛ಶࠊ࡛ಶࡢ㑇ఏᏊࢆ ࣆࢵࢡࢵࣉࡋࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡢ୰ࡣࣄ࣮ࢺ ࢩࣙࢵࢡࢱࣥࣃࢡ㉁ࠊ࣍ࣝࣔࣥࠊィ㛵㐃ࡢ㑇ఏᏊ ࡀྵࡲࢀ࡚࠸ࡓࠋࡉࡽࠊࠥࢆゎᯒࡋࠊ ࡢࡳ࡛Ⓨ⌧㔞ࡀ㧗ࡃࠊ㸦ࡶࡋࡃࡣపࡃ㸧࡛ࠥ ࡣ࠸ࡎࢀࡶⓎ⌧㔞ࡀప࠸㸦ࡶࡋࡃࡣ㧗࠸㸧㑇ఏᏊࢆ ಶྠᐃࡋࡓࠋ࠾ࡑࡽࡃࡇࢀࡽࡢ㑇ఏᏊࡢ୰ࡣఇ ╀ㄏᑟ㛵㐃ࡋࠊ%P753$ࡢάᛶࢩࢢࢼࣝࡢୗὶ ࡛ᶵ⬟ࡍࡿ㑇ఏᏊࡀྵࡲࢀ࡚࠸ࡿ⪃࠼ࡽࢀࡿࠋḟ ࠊࡢ⻈ࡢ⬻Ѹ㣗㐨ୗ⚄⤒⠇࡛ಶࡢ 㑇ఏᏊࢆࣆࢵࢡࢵࣉࡋࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡢ୰ ࡣࣄ࣮ࢺࢩࣙࢵࢡࢱࣥࣃࢡ㉁ࠊ࣍ࣝࣔࣥ㛵㐃ࡢ㑇ఏ Ꮚࡀྵࡲࢀ࡚࠸ࡓࠋ⯆῝࠸ࡇࠊࡢ ゎ ᯒ ࡛ ᕪ ࡢ ࠶ ࡗ ࡓ ಶ ࡢ 㑇 ఏ Ꮚ ࡢ ෆ ࠊ ಶ ࡢ㑇ఏᏊࡀ࡛ᕪࡢ࠶ࡗࡓ ಶࡢ㑇ఏᏊ㔜」ࡋ࡚࠸ࡓࠋࡇࡢࡇࡣࠊఇ╀ㄏᑟ ࡣࣛࣇࢫࢸ࣮ࢪ⯡ࢆ㏻ࡋ࡚ࡢ㑇ఏᏊⓎ⌧ࡢ㐪 ࠸ࡀ㔜せ࡛࠶ࡿࡇࢆ♧ࡋ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿࠋࡉ ࡽࠊᖺᗘࡢᨭࡼࡾࠊఇ╀ᛶࡢ㐪࠸ࡼࡾ ඹ㏻ࡢᣲືࢆ♧ࡍ㑇ఏᏊࢆྠᐃࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡾࠊఇ╀ㄏᑟ㛵ࢃࡿ⎔ቃ ᗘሗࡢ ཷᐜࡽ'+ࡢᨺฟไᚚ⧅ࡀࡿྍ⬟ᛶࡢ࠶ࡿ㑇ఏ Ꮚࡀከᩘࣆࢵࢡࢵࣉࡉࢀࡓࠋࡑࡢᚋࠊ573&5ゎᯒ ࣜࣝࢱ࣒3&5ゎᯒࡼࡾࠊ⯆῝࠸㑇ఏᏊࡢ ⬇ᮇࢆ㏻ࡋࡓⓎ⌧ࡸ⻈ᮇࡢ⬻Ѹ㣗㐨ୗ⚄⤒⠇࡛ࡢⓎ ⌧ࢆ☜ㄆࡋࡓࠋࡑࡋ࡚⌧ᅾࠊࡇࢀࡽࡢఇ╀ㄏᑟ㛵㐃 ೃ⿵㑇ఏᏊࡣࠊつᶍࢤࣀ࣒⦅㞟ᢏ⾡ࡢ7$/(1ࡸ ࢺࣛࣥࢫ࣏ࢰࣥࢆ⏝ࡋࡓᙧ㉁㌿࢝ࢥࢆసฟࡋࠊ ḟୡ௦ࡢఇ╀ᛶ࠼ࡿᙳ㡪ࢆㄪᰝࡍࡿࡶࠊ ࡑࢀࡽࡢಶయ࠾ࡅࡿ'+㔞࠼ࡿᙳ㡪ࡢㄪᰝࢆ ⾜࡞ࡗ࡚࠸ࡿࠋ࠶ࡿ㑇ఏᏊ㛵ࡋ࡚ࡣࠊẕ⻍ࡢ⬇ᮇ ࡛ࡢΥ࡛ࡢⓎ⌧ࡀΥẚ࡚᭷ព㧗ࡃࠊ 7$/(1ࡼࡿࣀࢵࢡ࢘ࢺ࢝ࢥ࡛ࡣḟୡ௦༸࠾ ࠸࡚ఇ╀ᛶࡢ㏫㌿ࡀほᐹࡉࢀࡓࠋ⌧ᅾࠊࡇࡢ㑇ఏᏊ ࡢᶵ⬟ゎᯒࢆ㔜Ⅼⓗ㐍ࡵࡿࡶࠊᮏᨭࡼ ࡗ࡚ᚓࡽࢀࡓ▱ぢࢆࡶࠊ࢝ࢥࡢ⬇ఇ╀ㄏᑟ ぢࡽࢀࡿⓎ⏕ྍረᛶ࠾ࡅࡿ⎔ቃሗࡢಖᏑ࣭グ᠈ ࡢศᏊᐇయࡢゎ᫂ࢆ┠ᣦࡋ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ሷぢ㑥༤⺋⣒࣭᪻ࣂ࢜ࢸࢵࢡ - 369 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࢿࢠᒓࣂ࢜ࣜࢯ࣮ࢫࢆ⏝࠸ࡓ࣑࢜ࢡࢫ⤫ྜゎᯒࡢࢱ࣐ࢿࢠ⫱✀ࡢᛂ⏝ ᇶ┙◊✲㸦㹀㸧 ᇳ⾜ ṇ⩏࣭ᒣཱྀᏛ㎰Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࢱ࣐ࢿࢠ㸦Allium cepa㸧ࡣࢤࣀ࣒ࢧࢬࡀ㠀ᖖࡁ ࡃ㸦15 Gbp㸧 㸪DNA ࣐࣮࣮࢝ࡢ㛤Ⓨ, 㐃㙐ᆅᅗࡢᵓ⠏, 㐃㙐ᆅᅗᰁⰍయࡢᑐᛂࡅ➼ࡢసᴗᅔ㞴ࢆక࠺ ࡓࡵ, ࡑࢀࢆඞ᭹ࡍࡿ┠ⓗ࡛␗✀㛫㞧ᰁⰍయಸ ຍࢆ⤌ࡳྜࢃࡏ࡚సฟࡋࡓࢩࣕࣟࢵࢺ㸦ศ⌫ᛶࢱ࣐ ࢿࢠ㸧⏤᮶༢୍␗✀ᰁⰍయࢆῧຍࡋࡓࢿࢠ⣔⤫ࢩࣜ ࣮ࢬ㸦ῧຍ⣔⤫ࢩ࣮ࣜࢬ㸧➼ࡢࣜࢯ࣮ࢫࡀ㛤Ⓨࡉࢀ ࡚ࡁࡓ㸬୍᪉࡛㸪ࡎࡉ DNA ◊㐃ᦠࡋ࡚ Allium Transcriptome Database㸦Allium TDB㸧ࡢ㛤Ⓨࡶ⾜ࡗ࡚ ࠾ࡾ㸪ࡇࡢ DB ᱁⣡ࡉࢀ࡚࠸ࡿྛ✀ࢿࢠᒓ✀ࡢࢺ ࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢹ࣮ࢱࡢ㓄ิẚ㍑ゎᯒࢆ⾜࠸㸪 ✀㛫ከᆺࢆ♧ࡍ⭾࡞ᩘࡢ DNA ࣐࣮࣮࢝ࢆᚓ࡚࠸ ࡿ㸬ࡋࡓࡀࡗ࡚㸪ࢩࣕࣟࢵࢺࢿࢠࡢ㛫࡛ᚓࡽࢀ࡚ ࠸ࡿ DNA ࣐࣮࣮࢝ࢆ༢୍␗✀ᰁⰍయῧຍ⣔⤫ࢩࣜ ࣮ࢬ㸦FF+1A-FF+8A㸧ࡼࡿከᆺゎᯒ㐺⏝ࡋ㸪ࡑࢀ ࡽࡀᗙࡍࡿࢩࣕࣟࢵࢺᰁⰍయࢆỴᐃࡍࡿࡇࡶྍ ⬟࡛࠶ࡿ㸬ࡑࡇ࡛㸪ᮏ◊✲࡛ࡣ㸪ྠࢩ࣮ࣜࢬࡢ᰿㒊 ࡽ RNA-Sequencing ࢹ࣮ࢱࢆ㞟ࡋ㸪ࢩࣕࣟࢵࢺ㫣 ⱼ㒊ࡢ unigene ࢭࢵࢺࢆࣜࣇࣞࣥࢫࡋ࡚࣐ࢵࣆ ࣥࢢࢆ⾜࡞࠺ࡇࡼࡾ㸪ࢿࢠ⏤᮶ࡢ㑇ఏᏊ㆑ู ࡛ࡁࡿࢩࣕࣟࢵࢺ㑇ఏᏊࡢሗࢆᩚ⌮ࡋ㸪ࡑࢀࢆᇶ ࡋ࡚ྠ᳜≀ࡢᰁⰍయ࣐࣮࣮࢝ྲྀᚓࢆඛࡎࡣヨࡳࡓ㸬 㸺ᨭࡢෆᐜ㸼 ࢩࣕࣟࢵࢺࡢᰁⰍయࢆῧຍࡋࡓࢿࢠῧຍ⣔⤫ࢩ࣮ࣜ ࢬࡢ᰿㒊࠾ࡅࡿ RNA-seq ゎᯒ㸦30 ᳨య㸧 㸸㸯᳨య ᙜࡾࡢ࣮ࣜࢻᩘࢆ 2,000 ࣮ࣜࢻタᐃࡋ㸪㸯㸮✀㢮 ࡢ᳜≀య㸦㸶✀㢮ࡢῧຍ⣔⤫㸰✀㢮ࡢ୧ぶ✀㸧 㹖 㸱 㸻㸱㸮᳨యࡢゎᯒᚲせ࡞࣮ࣜࢻᩘࡣ 20M 㹖 30 = 600M㸦6 ൨㸧࣮ࣜࢻ࡞ࡾ㸪ࡇࡢ 600M ࣮ࣜ ࢻࡣཷクゎᯒ࡛⏝ࡉࢀࡓ HiSeq ࢩ࣮ࢣࣥࢧ࣮࡛ࡢ 1 ࣇ࣮ࣟࢭࣝゎᯒ 1-2 ᅇศ㸦8 ࣮ࣞࣥ୰ 1-2 ࣮ࣞࣥ㸧 ࢆ࠼ࡤ㐩ᡂ࡛ࡁࡿᩘ್㸦100PE ࣛࣥࡢሙྜ㸧࡛࠶ ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ྛ⣔⤫ࡘࡁ 3 ࢧࣥࣉࣝࢆ⏝࠸, ྛࢧࣥࣉࣝ 20M reads ࡢ㓄ิሗࢆ㞟ࡋࡓ. ᚓࡽࢀࡓ㓄ิሗࡣ, ᖺᗘᵓ⠏ࡋࡓࢩࣕࣟࢵࢺ㫣ⱼ㒊ࡢ unigene ࢭࢵࢺ ࢆࣜࣇࣞࣥࢫࡋ࡚࣐ࢵࣉࡋࡓᚋ㸪RPKM㸦reads per kilobase of exon per million mapped sequence reads㸧 ್ࢆ⟬ฟࡍࡿࡇࡼࡾ㞟ィࡋ, ᩚഛࢆ㐍ࡵ࡚࠸ࡿ ࢿࢠᒓࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢹ࣮ࢱ࣮࣋ࢫ 㸦AlliumTDB㸧ሗࢆຍ࠼ࡿࡇࡼࡾẚ㍑ゎᯒ, ⤫ྜ࣑࢜ࢡࢫゎᯒᛂ⏝ࡍࡿࡓࡵࡢᇶ┙ሗࡋ࡚ ᩚഛࡋࡓ㸬RPKM ࢹ࣮ࢱᩚ⌮ࡢ⤖ᯝ, ῧຍࡋࡓࢩࣕ ࣟࢵࢺᰁⰍయ⏤᮶ࡍࡿ⪃࠼ࡽࢀࡿࢩࣕࣟࢵࢺࡢ Ⓨ⌧㑇ఏᏊࡢሗࡸ㸪ࢩࣕࣟࢵࢺᰁⰍయࡢῧຍࡼ ࡾⓎ⌧ㄏᑟࢀࡓ⪃࠼ࡽࢀࡿࢿࢠ㑇ఏᏊࡢሗࢆ ᚓࡿࡇࡀ࡛ࡁࡓ㸬ຍ࠼࡚㸪RNA-Sequencing ࡛ᚓࡽ ࢀࡓࢩࣕࣟࢵࢺࢿࢠࡢ㛫ࡢ㓄ิከᆺࡢᙳ㡪࡛㸪ࢩ ࣕࣟࢵࢺ⏤᮶ࡢ㑇ఏᏊࡢࡳࡀ࣐ࢵࣉࡉࢀࡿ㸦ࢿࢠぶ ᰴࡢ RPKM ್ࡀ 0 ࡞ࡿ㸧unigene ࢆ᳨⣴ࡋࡓࡇࢁ㸪 ᰿㒊࡛ 10,197 ࡢᑐ㇟㓄ิࡀࡑࢀࡒࢀᚓࡽࢀࡓ㸬ࡉࡽ 㸪≉ᐃࡢᰁⰍయ⣔⤫ࡢࡳ࡛ࢩࣕࣟࢵࢺ㏆࠸ RPKM ್ࡀᚓࡽࢀࡓ unigene ࡢᩘࢆ࢝࢘ࣥࢺࡍࡿ 5,308 ࡞ࡗࡓ㸬ࡑࢀࡽࡢᰁⰍయẖࡢෆヂࡣ௨ୗࡢ㏻ ࡾ࡞ࡗࡓ㸹1A 803, 2A 830, 3A 799, 4A 594, 5A 603, 6A 622, 7A 568, 8A 489. ࡲࡓ㸪ࡇࢀࡽࡢ୰ࡣ㸪ࡑࡢ ࡢ㸰ჾᐁ㸦ⴥ㌟ⴥ㠧㸧ࡢゎᯒࡢ࠸ࡎࢀ࠾࠸࡚ ࡶ᳨ฟࡉࢀ࡚࠸ࡿඹ㏻ unigene ࡶከᩘᚓࡽࢀ࡚࠾ࡾ㸪 ࡑࢀࡽࡀ᭱ࡶ☜ᗘࡢ㧗࠸ᰁⰍయ≉␗ⓗ unigene ࡛࠶ ࡾ㸪࣐࣮࣮࢝ࡋ࡚ඃඛⓗά⏝࡛ࡁࡿࡇࡀ♧၀ ࡉࢀࡓ㸬௨ୖࡼࡾ㸪ῧຍ⣔⤫ࢩ࣮ࣜࢬࢆ⏝࠸ࡓࢺࣛ ࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࡼࡾ㸪ᰁⰍయ≉␗ⓗ࣐࣮࢝ ࣮ࡢ㔞ྲྀᚓࡀ㐩ᡂࡉࢀࡓ㸬⌧ᅾ㸪ࢱ࣐ࢿࢠಸຍ༙ ᩘయ㸦DH㸧ࢩࣕࣟࢵࢺ DH ࡢࡅྜࢃࡏࡼࡾస ฟࡉࢀࡓ F2 ศ㞳㞟ᅋࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢹ࣮ࢱ ࢆᇶࡋࡓ㑇ఏᏊᆺሗࡶᚓࡽࢀጞࡵ࡚࠾ࡾ㸪ᮏ◊ ✲ࡼࡾᚓࡽࢀࡓሗࡣ㔞ࡢ SNP ࣐࣮࣮࢝➼ࢆໟ ྵࡍࡿ㉸㧗ᐦᗘ㐃㙐ᆅᅗࢆ≀⌮ᰁⰍయࡾศࡅࡿ 㝿ຠຊࢆⓎࡍࡿᛮࢃࢀࡿ㸬 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᡃࠎࡣ㸪㐃㙐ᆅᅗస〇ࡸ QTL ゎᯒྥࡅࡓ࣓ࢱ࣎ࣟ ࣮࣒ሗࡢ㞟ࢆ㐍ࡵ࡚࠸ࡿୖグ F2 ಶయᑐࡋ࡚ࢺ ࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ⾜࠸㸪ᚓࡽࢀࡓ㓄ิሗ ࡢ SNP ࣃࢱ࣮ࣥࢆゎᯒࡍࡿࡇࡼࡾ㸪ࢱ࣐ࢿࢠ⏤ ᮶ࢩࣕࣟࢵࢺ⏤᮶ࡢ㌿⏘≀ࢆ༊ูࡋ㸪ࡑࡢሗ ࢆᇶ㑇ఏᏊᆺሗࢆ㞟ࡋጞࡵ࡚࠸ࡿ㸬30 ಶయࢆ ⏝࠸ࡓࢸࢫࢺゎᯒࡼࡾᚓࡽࢀࡓ RNA-seq ࡢ㓄ิሗ ࢆࢩࣕࣟࢵࢺ㫣ⱼ㒊ࡢ unigene ࢭࢵࢺࢆࣜࣇࣞࣥ ࢫࡋ࡚࣐ࢵࣉࡋ㸪ሷᇶከᆺሗࢆ㞟ᩚ⌮ࡍࡿࡇ ࡼࡾ㸪⣙ 5,000 ࡢ unigene ࡘ࠸࡚㑇ఏᏊᆺሗ ࢆᚓࡿࡇࡀ࡛ࡁ࡚࠸ࡿ㸬ࡇࡢሗࢆᩚഛ୰ࡢ㑇ఏ Ꮚᆅᅗሗ⤫ྜࡍࡿࡇ࡛㸪࣓ࢱ࣮࣒࣎ࣟ QTL ࡸ ࢤࣀ࣒㓄ิゎᯒྥࡅࡓ㧗ᐦᗘ࣐࣮࣮࢝ࡢᩚഛ➼ ά⏝࡛ࡁࡿࡇࡀ♧၀ࡉࢀ࡚࠾ࡾ㸪ᮏ◊✲࡛ᚓࡽࢀ ࡓ࣐࣮࣮࢝ሗ↷ྜࡍࡿࡇ࡛▷ᮇ㛫㉸㧗ᐦᗘ ᰁⰍయᆅᅗࢆᵓ⠏࡛ࡁࡿྍ⬟ᛶࡀษࡾ㛤ࢀࡓ࠸ ࠼ࡼ࠺㸬 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋࠋ - 370 - බເᨭㄢ㢟㸸2011㹼2012 ᖺᗘ ᅵተᚤ⏕≀ࡢᆒ➼ᗘホ౯ᡭἲࡢ㛤Ⓨ ᇶ┙◊✲㸦㹀㸧 ಙ⃰ ༟㑻࣭㎰ᴗ࣭㣗ရ⏘ᴗᢏ⾡⥲ྜ◊✲ᶵᵓᾏ㐨㎰ᴗ◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ᅵተᚤ⏕≀ࡣ✀ᩘࠊಶయᩘࡀከ࠸ୖ㞴ᇵ㣴ᛶ࡛ ࠶ࡿࡓࡵ◊✲ࡀ㐜ࢀ࡚࠸ࡿࠋࡑࡇ࡛㉸୪ิᆺࢩ࣮ ࢡ࢚ࣥࢧ࣮ࢆ⏝࠸ࠊᅵተࡽᢳฟࡋࡓ DNA/RNA ࢆᑐ㇟ᶵ⬟ᛶ㑇ఏᏊࡢࣥࣉࣜࢥࣥࢩ࣮ࢡ࢚ࣥ ࢫゎᯒࢆ⾜࠸ࠊᶵ⬟ᛶ㑇ఏᏊࡢከᵝᛶホ౯ࢆ⾜࠺ࠋ ࡉࡽᅵተࡽᢳฟࡋࡓ DNA ࢆ┤᥋ࢩ࣮ࢡ࢚ࣥ ࢫࡋࠊࣥࣉࣜࢥࣥࢩ࣮ࢡ࢚ࣥࢫゎᯒࡼࡾᚓࡽ ࢀࡿࢹ࣮ࢱྜࢃࡏࡿࡇ࡛ᆒ➼ᗘࡼࡿホ౯ࢆ ⾜࠺ࠋࡇ࠺ࡋ࡚ᚓࡽࢀࡓᡂᯝࢆඖࠊᶵ⬟ᛶ㑇ఏ Ꮚࡢᆒ➼ᗘ⫧Ỻᗘࡸࣂ࢜ࢪ࢙࢜ࢣ࣑࢝ࣝࣉࣟ ࢭࢫࡢ㛵㐃ᛶࢆ᫂ࡽࡋ࡚᪂ࡓ࡞◊✲ࡢ᪉ྥ ᛶࢆᡴࡕฟࡍࠋ ࣉࣜࢥࣥࢩ࣮ࢡ࢚ࣥࢫ⤖ᯝࡢゎᯒࠊࢹ࣮ࢱ࣮࣋ ࢫࠊྛ㑇ఏᏊ㛫ࡢ㐃㙐ゎᯒࢆ⾜࠺ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㑅ᐃࡍࡿ㑇ఏᏊࢆ⏝࠸ࡓሗゎᯒࡢྍ⬟ᛶࡘ࠸࡚ ㆟ㄽࢆ㐍ࡵࠊࡑࢀᇶ࡙ࡃᅵተᚤ⏕≀ࡢゎᯒࢆ⾜ࡗ ࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ከ✀ከᵝ࡞ᚤ⏕≀㑇ఏᏊࡽᵓᡂࡉࢀࡿ⎔ቃᶵ⬟ࢆ ゎᯒࡍࡿᡭἲࡋ࡚ά⏝ྍ⬟࡛࠶ࡿࡇࡀ⪃࠼ࡽࢀ ࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 㸺ᨭࡢෆᐜ㸼 ࣥࣉࣜࢥࣥࢩ࣮ࢡ࢚ࣥࢫࡼࡿ⥙⨶ⓗ࡞ᐃᛶ ゎᯒࡢᑟධࢆヨࡳ࡚࠸ࡿࠋึᖺᗘᙜࡓࡿᮏᖺ ࡣ⣔⤫Ꮫⓗᣦᶆ㑇ఏᏊࡸᆅ⌫⎔ቃᐦ᥋㛵 ࡋ࡚࠸ࡿ⪃࠼ࡽࢀ࡚࠸ࡿᅵተᚤ⏕≀ࡼࡿࣂ ࢜ࢪ࢙࢜ࢣ࣑࢝ࣝࣉࣟࢭࢫࡢ࠺ࡕᮏゎᯒ⏝ ࠸࠺ࡿ㑇ఏᏊࢆྛ ⛬ᗘ㑅ᢤࡋࠊྜィ 㑇ఏᏊ ࡢゎᯒࢆ⾜࠺ࠋࡲࡓᑐ㇟ࡍࡿᅵተࡣࠊ୍⯡ⓗ ࡞⟶⌮ࡀࡉࢀࡓᅡሙຍ࠼ࠊ᭷ᶵ㎰ἲࡀࡉࢀ ࡓᅡሙࡸ⣙㸯㸮㸮ᖺ㛫␗࡞ࡿ⫧⟶⌮ࢆ⾜ࡗ࡚ ࡁࡓᅡሙࠊᙉຊ࡞ ᬮ࢞ࢫ࡛࠶ࡿ 12 ࢞ࢫࢆ 㔞ᨺฟࡍࡿᅡሙ࡞㸯㸮ᆅⅬ⛬ᗘࢆணᐃࡋ ࡚࠸ࡿࠋḟᖺᗘ௨㝆ࠊゎᯒᑐ㇟ࡍࡿ㑇ఏᏊᩘ ࢆቑࡸࡍணᐃ࡛࠶ࡿࠋලయⓗࡣࡇࢀࡽࡢࣥ - 371 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࣑ࢺࢤࣀ࣒ゎᯒ᰾࣐࣮࣮࢝ࢱࣆࣥࢢࡼࡿ᪥ᮏ⏘ࣁࣈᒓ ✀ࡢ㑇ఏⓗ㞟ᅋᵓ㐀ࡢ◊✲ ᇶ┙◊✲㸦㹁㸧 ᰘ⏣ ᘯ⣖࣭ᕞᏛ⏕య㜵ᚚ་Ꮫ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ་Ꮫⓗࡶ⏕ែᏛⓗࡶࡲࡓ⸆ࢩ࣮ࢬࡋ࡚ࡶ 㔜せ࡞ẘ⏕≀࡛࠶ࡿࣁࣈࡢ㑇ఏⓗከᵝᛶࢆໟᣓⓗ ⌮ゎࡍࡿࡇࢆ┠ᣦࡋ࡚ࠊ᪥ᮏ⏘ࣁࣈᒓ 3 ✀㸦ࣁࣈ 㸦 Protobothrops flavoviridis 㸧ࠊ ࢺ ࢝ ࣛ ࣁ ࣈ 㸦 P. tokarensis㸧ࠊࢧ࢟ࢩ࣐ࣁࣈ㸦P. elegans㸧㸧ࡘ࠸࡚ ⏕ᜥᇦᇦࢃࡓࡿつᶍ DNA ゎᯒࢆ㐍ࡵࡿࠋලయ ⓗࡣ௨ୗࡢ㸰ࡘࡢィ⏬ࡽ࡞ࡿࠋ ձᓥ㞟ᅋ㛫ࡢ㑇ఏⓗศࡢᗘྜ࠸ࢆࠊ࣑ࢺࢥࣥࢻࣜ ࢤࣀ࣒ཬࡧ࣐ࢡࣟࢧࢸࣛࢺ࣐࣮࣮࢝ࡢከᵝᛶ ࡼࡾ᥎ᐃࡋࠊࣁࣈᒓ 3 ✀ࡢ⣔⤫ᆅ⌮ࢆໟᣓⓗゎ ᫂ࡍࡿࠋ࠶ࢃࡏ࡚⌧⾜ࡢ 3 ✀ศ㢮ࡢጇᙜᛶࢆ᳨ウࡍ ࡿࠋ ղୖ࡛ྠᐃࡉࢀࡿከᆺᛶࡢ㧗࠸㡿ᇦࠊ࣐࣮࣮࢝ᑐ ㇟ࢆ⤠ࡗ࡚ࠊྛᓥ㞟ᅋෆࡢ㑇ఏⓗከᵝᛶࢆ᳨ฟࡋࠊ 㑇ఏᏛⓗ࡞㞟ᅋᵓ㐀ࡢ᥎ᐃࢆ⾜࠺ࠋ㑇ఏᏊὶືࡢṔ ྐࢆ⌧ࡋࠊࣁࣈᒓࡢ⌧ᅾࡢศᕸᇦࡀᙧᡂࡉࢀࡿ㐣 ⛬ࢆ༡すㅖᓥࡢᆅྐ↷ࡽࡋ࡚ᵓ⠏ࡍࡿࠋࡉࡽࠊ 㞟ᅋ㑇ఏᏛⓗࣃ࣓࣮ࣛࢱࡽྛ㞟ᅋࡢ㑇ఏⓗᗘ ࢆ᳨ウࡋࠊ✀ࡢಖ㈨ࡍࡿࠋ 㸦ୗᅗ㸧ࠋ⨾⩌ᓥἈ⦖ㅖᓥࡢᆅ⌮Ꮫⓗ࡞ศ᩿ࡣ 150 ᖺ๓᥎ᐃࡉࢀ࡚࠾ࡾࠊࣁࣈࡢ♽ඛ㞟ᅋࡣᆅ ⌮Ꮫⓗ࡞ศ᩿ඛ❧ࡗ࡚ࠊ⨾ࢡ࣮ࣞࢻ⌰⌫ࢡࣞ ࣮ࢻ㑇ఏⓗศࡋ࡚࠸ࡓࡇࡀ♧၀ࡉࢀࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࣁࣈ㸦Ἀ⦖ᮏᓥ⏘㸧ࡢ WGS ࢩ࣮ࢡ࢚ࣥࢩࣥࢢ ᪂つࢭࣥࣈࣜࡼࡿࢻࣛࣇࢺ㓄ิసᡂࠋࢺ࢝ࣛࣁ ࣈ࠾ࡼࡧࢧ࢟ࢩ࣐ࣁࣈࡢ WGS ࢩ࣮ࢡ࢚ࣥࢩࣥࢢ Ἀ⦖ࣁࣈࢤࣀ࣒ࢻࣛࣇࢺ㓄ิࡢ࣐ࢵࣆࣥࢢࡼࡿ ኚ␗ゎᯒࠋࣁࣈࡢẘ⭢㸦ᡂయ࠾ࡼࡧᗂయ㸧ࠊẘ⭢ࠊ ⮅⮚ࠊ⫢⮚ࠊ༸ᕢࡽࡢ RNA ࢩ࣮ࢡ࢚ࣥࢩࣥࢢࠋ ᨭࢆ࠾㢪࠸ࡋ࡚࠸ࡿࣁࣈᒓ 3 ✀ࡢࢤࣀ࣒㓄ิ ࡣᘬࡁ⥆ࡁゎᯒ୰࡛ࢹ࣮ࢱࡢྲྀᚓࡣ⮳ࡗ࡚࠸࡞࠸ ࡀࠊࡇࢀࡀᡂࡍࢀࡤࠊ࣐ࢡࣟࢧࢸࣛࢺࢆጞࡵ ࡍࡿ᰾㑇ఏᏊ࣐࣮࣮࢝ࡢ㞟ࡀ⏝ព࡞ࡾࠊ㑇ఏ ⓗከᵝᛶࡢ⌮ゎࡣ㣕㌍ⓗ㐍ࡴᮇᚅࡉࢀࡿࠋ≉ ከᆺᛶࡢ㧗࠸࣐ࢡࣟࢧࢸࣛࢺ࣐࣮࣮࢝ࡀᩚഛࡉ ࢀࡿࡇ࡛㞟ᅋෆࡢヲ⣽࡞ከᵝᛶゎᯒࡀྍ⬟࡞ࡿ ᮇᚅࡉࢀࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⨾ᓥࠊຍィ࿅㯞ᓥࠋᚨஅᓥࠊἈ⦖ᮏᓥࠊఀᖹ ᒇᓥࠊఀỤᓥࠊஂ⡿ᓥࠊΏᩜᓥࡢ 6 ᓥࡢࣁࣈྛ 4 ࡲࡓࡣ 5 ಶయࠊᑠᐆᓥࠊᐆᓥࡢࢺ࢝ࣛࣁࣈྛ 4 ಶయࠊ ࠾ࡼࡧ▼ᇉᓥࠊす⾲ᓥࡢࢧ࢟ࢩ࣐ࣁࣈྛ 1 ಶయࠊཬ ࡧእ⩌ࡋ࡚ࣄ࣓ࣁࣈ㸦Ovophis okinavensis㸧1 ಶ యࠊィ 44 ಶయࡢ࣑ࢺࢤࣀ࣒㓄ิࢆỴᐃࡋ᭱ᑬἲࡼ ࡿゎᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࣁࣈࡣ, 㒊ಶయ⩌㸦⨾ ᓥ, ຍィ࿅㯞ᓥࠊᚨஅᓥ㸧ࡽ࡞ࡿ⨾ࢡ࣮ࣞࢻࠊ ༡㒊ಶయ⩌㸦Ἀ⦖ᓥ, ఀᖹᒇᓥ, ఀỤᓥࠊஂ⡿ᓥࠊΏ ᩜᓥ㸧ࡽ࡞ࡿ⌰⌫ࢡ࣮ࣞࢻࡢ 2 ⩌㑇ఏⓗศ ࡋ࡚࠸ࡿࡇࡀศࡗࡓ㸦ᅗ 1㸧 ࠋࡲࡓ, ࢺ࢝ࣛࣁ ࣈࡣᐆᓥࠊᑠᐆᓥࡶࠊ⨾ࢡ࣮ࣞࢻໟ ྵࡉࢀࡓ. ௨ୖࡢࡇࡽࠊ⌧ᅾࣁࣈࡉࢀࡿ✀ࡢᓥ 㞟ᅋ㛫ࡢ㑇ఏⓗከᵝᛶࡣࡁࡃࠊ⌧ᅾู✀ࡉࢀࡿ ࢺ࢝ࣛࣁࣈࡣࡑࡢከᵝᛶࡢ୰ࡲࡿࡓࡵࠊ ࢺ࢝ࣛࣁࣈࡢࡳࢆ⊂❧✀ࡍࡿࡇࡢጇᙜᛶࡣప࠸ ⪃࠼ࡽࢀࡓࠋ୍᪉ࠊࢧ࢟ࢩ࣐ࣁࣈࡘ࠸࡚ࡣࡑࡢ ༢⣔⤫ᛶࡀ☜ㄆࡉࢀࡓࠋࡉࡽྜయࢩ࣑࣮ࣗࣞࢩࣙ ࣥࡼࡾศᒱᖺ௦ࢆ᥎ᐃࡋࡓࡇࢁࠊ⨾ࢡ࣮ࣞࢻ ⌰⌫ࢡ࣮ࣞࢻࡢศᒱࡣࠊ529 ᖺ๓᥎ᐃࡉࢀࡓ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ẘ⭢ࢆጞࡵࡍࡿࣁࣈ⤌⧊ࡽࡢ RNA-seq ࡢࢹ ࣮ࢱࢆࠊࢤࣀ࣒ᨭࡼࡗ࡚ྲྀᚓࡍࡿࡇࡀ࡛ࡁࡓࠋ ࡇࢀࢆࢭࣥࣈࣝࡋࡓ㌿⏘≀ሗࠊࡇࢀࡲ࡛ ᚓࡽࢀ࡚࠸ࡓࣁࣈࢤࣀ࣒ࡢࢻࣛࣇࢺ㓄ิࢆẚ㍑ࡍࡿ ࡇ࡛ࠊࣁࣈࡢẘᾮ㛵㐃㑇ఏᏊࡢ㓄ิࡀ 329 ಶྠᐃ ࡉࢀࡓࠋࡇࢀࡽࡢ㑇ఏᏊ㓄ิሗࠊᮏ⛉◊ㄢ㢟࡛ ࡍ࡛㞟῭ࡳࡢከᩘࡢࣁࣈ㞟ᅋ⏤᮶ࡢࢧࣥࣉࣝࢆ ⏝࠸ࡿࡇ࡛ࠊẘᾮ㛵㐃㑇ఏᏊ⩌ࡢ㑇ఏⓗከᵝᛶࡢ ໟᣓⓗ࡞ゎᯒࢆ⾜࠺ࡇࡀྍ⬟࡞ࡗࡓࠋࡇࢀࡼ ࡾࠊࣁࣈẘᾮᵓᡂᡂศࡢ࠸ࡃࡘࡢ㑇ఏᏊ࡛ሗ࿌ࡉ ࢀ࡚࠸ࡿຍ㏿㐍ࡢ୍⯡ᛶࡸࠊࡑࡢ࣓࢝ࢽࢬ࣒㏕ ࡿ◊✲⧅ࡀࡿᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Shibata et al. Genetic divergence of mitochondria genome sequence of the Habu viper, Protobothrops among Japanese subtropical islands.㸦ᢞ✏୰㸧 - 372 - බເᨭㄢ㢟㸸2011 ᖺᗘࠊ2012 ᖺᗘ ᳜≀࠾ࡅࡿ࢚ࣆࢤࣀ࣒ࢆࡋࡓඃຎᛶⓎ⌧ไᚚᶵᵓࡢゎ᫂ ᭱ඛ➃࣭ḟୡ௦◊✲㛤Ⓨᨭࣉࣟࢢ࣒ࣛ ᰘ ༤ྐ࣭ዉⰋඛ➃⛉Ꮫᢏ⾡Ꮫ㝔Ꮫࣂ࢜ࢧ࢚ࣥࢫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ᭷ᛶ⏕Ṫࡼࡗ࡚⏕ࡳฟࡉࢀࡿᏊᏞࡣࠊ୧ぶࡢᣢ ࡘᛶ㉁ࡢ࠸ࡎࢀ୍᪉ࡢࡳࢆཷࡅ⥅ࡄሙྜࡀከࡃ▱ ࡽࢀ࡚࠸ࡿࠋ࣓ࣥࢹࣝࡢࠕඃᛶࡢἲ๎ࠖࡋ࡚▱ࡽ ࢀࡿ㑇ఏ⌧㇟࡛࠶ࡿࡀࠊࡑࡢ࣓࢝ࢽࢬ࣒ࡣ᫂࡞Ⅼ ࡀከ࠸ࠋᡃࠎࡣࠊ᳜≀ࡢཷ⢊㛵ࢃࡿᅉᏊࡢ◊✲ࢆ ㏻ࡌ࡚ࠊࢤࣀ࣒ሷᇶ㓄ิࡢኚࢆక࠺ࡇ࡞ࡃᙧ㉁ ኚ␗ࡀ⏕ࡌࡿࡇ࡛ඃຎᛶࢆỴᐃࡍࡿ࠸࠺᪂ࡋ࠸ ඃຎᛶỴᐃᶵᵓࢆୡ⏺ඛ㥑ࡅ࡚᫂ࡽࡋࡓࠋᮏ ◊✲ㄢ㢟࡛ࡣࠊ᭱᪂ࡢࢤࣀ࣒ゎᯒᢏ⾡ࢆ㥑ࡋ࡚ࠊ ᳜≀࠾ࡅࡿࢤࣀ࣒ሷᇶ㓄ิࡢኚࢆకࢃ࡞࠸ඃຎ ᛶⓎ⌧ไᚚࡢᐇែࢆࢤࣀ࣒࣡ࢻ᫂ࡽࡍࡿ ࡶࠊࡑࡢศᏊᶵᵓゎ᫂ࢆ┠ᣦࡍࠋࡲࡓ✀ෆ㞧✀ ᙜ௦࡛≉␗ⓗ࡞㑇ఏᏊⓎ⌧ไᚚᶵᵓࡢᐇែࢆ᫂ࡽ ࡍࡿࠋࡉࡽᚤ㔞⤌⧊ࡽࡢ DNA ࣓ࢳࣝࣃࢱ࣮ ࣥࡢ⥙⨶ⓗゎᯒࢆ㏻ࡌ࡚ࠊ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞㑇 ఏᏊⓎ⌧ไᚚࡀᣦࡉࢀ࡚࠸ࡿⰼ⢊ࡢ⏕Ṫ⣽⬊ᰤ 㣴⣽⬊࠾ࡼࡧⵂࢱ࣮࣌ࢺ⣽⬊࠾ࡅࡿ⤌⧊≉␗ⓗ࡞ ࣓ࢳࣝࡢᐇែࢆ᫂ࡽࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ୍㑇ఏᏊከᆺ613ࢆ⪃៖ࡋࡓࣞࣝ≉␗ⓗ࡞ 㑇ఏᏊⓎ⌧ࣃࢱ࣮ࣥࡢ⥙⨶ⓗࡘᐃ㔞ⓗゎᯒ 613 ሗࢆᚓࡿࡓࡵࡢࢩࣟࢾࢼࢬࢼ✀ෆ⣔⤫ ࡢࣜࢩ࣮ࢡ࢚ࣥࢫ ࢩࣟࢾࢼࢬࢼ✀ෆ⣔⤫ࡢ VPDOO51$ ࢩ࣮ࢡ࢚ ࣥࢫ ࣂࢧࣝࣇࢺࢩࣙࢵࢺ࢞ࣥࢩ࣮ࢡ࢚ࣥࢫ ࡼࡿࢩࣟࢾࢼࢬࢼ✀ෆ㞧✀࠾ࡅࡿ '1$ ࣓ࢳࣝ㒊ࡢゎᯒ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ➹⪅ࡣࠊࢤࣀ࣒㓄ิࡀබ㛤ࡉࢀ࡚࠸࡞࠸ࢩࣟ ࢾࢼࢬࢼ✀ෆ⣔⤫ࡢࢤࣀ࣒㓄ิࢆࠊḟୡ௦ࢩ࣮ࢡ࢚ ࣥࢧ࣮ࢆ⏝࠸࡚ࣜࢩ࣮ࢡ࢚ࣥࢫࡋࠊ᪤බ㛤ࡉࢀ࡚ ࠸ࡿࢩࣟࢾࢼࢬࢼࡢࢤࣀ࣒ࢩ࣮ࢡ࢚ࣥࢫẚ㍑ࡋ ࡚ࠊ2 ⣔⤫㛫ࡢ SNP ሗࢆᚓࡓࠋࡑࡢ㝿ࠊSNP ゎᯒ ࢶ࣮ࣝࣃ࣓࣮ࣛࢱ᮲௳ࢆྫྷࡍࡿࡇ࡛ࠊ㧗⢭ᗘ ࡞ SNP ሗࢆᚓࡓࠋୖグ 2 ⣔⤫ࡇࢀࡽࢆࡅྜࢃ ࡏࡓ F1 㞧✀ࡢᐇ⏕ࡽ poly A RNA ࢆᢳฟࡋࠊḟୡ ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮࡛ࡑࡢ㑇ఏᏊⓎ⌧ࢆ⥙⨶ⓗゎᯒ ࡋࡓࠋࡑࡋ࡚ SNP ሗࢆ⏝࠸࡚୧ぶ⣔⤫࡛ඹⓎ⌧ ࢆ♧ࡍࡀࠊF1 㞧✀࡞ࡿ࠸ࡎࢀࡢぶ⏤᮶ࡍࡿ ᑐ❧㑇ఏᏊࡢࡳࡀⓎ⌧ࢆ♧ࡍ㑇ఏᏊࢆ᥈⣴ࡍࡿࡇ ࡼࡾࠊF1 㞧✀࠾࠸࡚∦ഃᑐ❧㑇ఏᏊⓎ⌧ࢆ♧ࡍ 㑇ఏᏊࢆ」ᩘྠᐃࡋࡓࠋࡲࡓ F1 㞧✀୧ぶ⣔⤫ࡢ small RNA ࡢ⥙⨶ⓗゎᯒࡶ⾜࠸ࠊඃຎᛶࡀࢺࣛࣥࢫ స⏝ᛶࡢపศᏊ RNA ࡼࡾㄪ⠇ࡉࢀ࡚࠸ࡿ᥎ ࡉࢀࡿᐇࢆぢฟࡋࡓࠋࡑࡇ࡛ F1 㞧✀୧ぶ⣔⤫ࡢ ᐇ⏕ࡽࢤࣀ࣒ DNA ࢆᢳฟࡋࠊࣂࢧࣝࣇࢺࢩ ࣙࢵࢺ࢞ࣥࢩ࣮ࢡ࢚ࣥࢫ౪ࡋ࡚ࠊDNA ࣓ࢳࣝࣃ ࢱ࣮ࣥࢆ⥙⨶ⓗ᳨ฟࡋࡓࠋF1 㞧✀୧ぶࡢ DNA ࣓ࢳࣝࣃࢱ࣮ࣥࡢ㐪࠸∦ഃᑐ❧㑇ఏᏊⓎ⌧ไᚚ ࡢ㛵ಀࢆㄪࡓࡀࠊ࣓ࢳࣝࣞ࣋ࣝ㑇ఏᏊⓎ⌧ ┦㛵㛵ಀࡣぢࡽࢀ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡾ✀ෆ⣔⤫㛫ࡢ 613 ሗࢆࢤࣀ࣒࣡ ࢻ㞟ࡍࡿࡇࡀฟ᮶ࡿࡼ࠺࡞ࡾࠊ) 㞧✀ ࠾ࡅࡿࣞࣝẖࡢඃຎⓎ⌧ࢆ⥙⨶ⓗㄪࡿ ࡀྍ⬟࡞ࡗࡓࠋࡑࡋ࡚ࠊඃᛶഃᑐ❧㑇ఏᏊ⏤᮶ ࡢపศᏊ 51$ ࡼࡾຎᛶഃᑐ❧㑇ఏᏊࡢⓎ⌧ࡀᢚ ไࡉࢀࡿ࠸࠺ඃຎᛶࣔࢹࣝࢆᥦၐࡍࡿࡇࡀ࡛ ࡁࡓࠋࡇࡢࡇࡣࠊᑐ❧㑇ఏᏊ㛫ࡢඃຎᛶ࠸࠺ 㔜せ࡞㑇ఏᏊⓎ⌧ไᚚᶵᵓࡢ⌮ゎࡘ࡞ࡀࡿ⪃ ࠼࡚࠸ࡿࠋࡲࡓᮏᨭࡼࡾࠊ) 㞧✀࠾ࡼࡧࡑࡢ ぶ⣔⤫࠾ࡅࡿ '1$ ࣓ࢳࣝࣃࢱ࣮ࣥࢆࢤࣀ࣒࣡ ࢻゎᯒࡍࡿࡇࡀฟ᮶ࡿࡼ࠺࡞ࡗࡓࠋࡇࢀ ࡼࡾࠊᑐ❧㑇ఏᏊ㛫ࡢ '1$ ࣓ࢳࣝㄏᑟᶵᵓࡸ ࡑࡢ⏕≀Ꮫⓗព⩏ࡘ࠸࡚ࡶ᫂ࡽ࡞ࡿ⪃࠼ ࡚࠸ࡿࠋᮏ◊✲ࡼࡗ࡚ᚓࡽࢀࡓ⤖ᯝࡣࠊ᳜≀ࡢ ඃຎᛶⓎ⌧㛵ࢃࡿ '1$ ࣓ࢳࣝᶵᵓࡢゎ᫂㈉ ⊩ࡍࡿࡤࡾ࡛࡞ࡃࠊື᳜≀ࢆྵࡵ࡚ᗈࡃ㑇ఏᏊ Ⓨ⌧ㄪ⠇ࡢ◊✲ᐤࡍࡿ⪃࠼ࡽࢀࡿࠋࡲࡓᙜ ヱ◊✲࡛ᚓࡽࢀࡓ ) 㞧✀ᙧᡂ࢚ࣆࢪ࢙ࢿࢸ ࢵࢡ࡞せᅉࡼࡾ㑇ఏᏊⓎ⌧ไᚚࡀ㉳ࡇࡿ㑇ఏᏊ ࢆ᫂ࡽࡋ࡚࠸ࡃࡇ࡛ࠊ᭷⏝ࣁࣈࣜࢵࢻస ≀ࡢసฟࡼࡿ⏕⏘ᶵ⬟ྥୖࡸࣂ࣐࢜ࢫᨵⰋ ࡘ࡞ࡀࡿࡀᮇᚅࡉࢀࡿࠋࡲࡓ㑇ఏᏊ⤌࠼ࢆ ࢃ࡞࠸᪂ࡋ࠸⫱✀ἲࡢ☜❧ࡶࡘ࡞ࡀࡿࡀᮇᚅ ࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᰘ ༤ྐ᳜≀ࡢ⏕㛗ㄪ⠇ 7DNDGD<* ๓ᕝ㞞ᙪࠊ⫱✀Ꮫ◊✲ 6KLED+ HWDO'*' ᰘ ༤ྐ㺂ᅜ㝿㧗➼◊✲ᡤ◊✲ࣉࣟࢪ࢙ࢡ ࢺࠕ⣽⬊ᒚṔᇶ࡙ࡃ᳜≀ࡢᙧែᙧᡂࠖሗ࿌᭩ S - 373 - බເᨭㄢ㢟㸸2012 ᖺᗘ 㑇ఏᛶ②ᛶᑐ㯞⑷ࡢ᪂つཎᅉ㑇ఏᏊྠᐃࡑࡢᶵ⬟ゎᯒ ᇶ┙◊✲㸦㹁㸧 ᔱᓮ ᬕ㞝࣭⮬་⛉Ꮫ་Ꮫ㒊 ෆ⛉Ꮫㅮᗙ ⚄⤒ෆ⛉Ꮫ㒊㛛 㸺⫼ᬒ࣭┠ⓗ㸼 㑇ఏᛶ②ᛶᑐ㯞⑷㸦HSP㸧ࡣ㸪⮫ᗋⓗ⦆ᚎ㐍⾜ ᛶࡢ୧ୗ⫥ࡢ②ᛶᑐ㯞⑷ࢆ୰᰾≧ࡋ㸪ࡑࡢ ᵝࠎ࡞≧ࢆྜేࡋ࠺ࡿೃ⩌࡛࠶ࡿ㸬㏆ᖺHSPࡢ 㑇ఏᏊᗙ࠶ࡿ࠸ࡣཎᅉ㑇ఏᏊࡀ㏆ᖺḟࠎゎ᫂ࡉࢀ ࡚ࡁ࡚࠸ࡿ㸬⌧ᅾࡲ࡛ࡢᡤ㸪ࡑࡢ㑇ఏᏊᗙࡣSPG1ࠥ 75ࡲ࡛ᰁⰍయୖ࣐ࢵࣆࣥࢢࡉࢀ࡚࠾ࡾ㸪ࡑࡢ࠺ࡕ ⣙60ಶవࡾࡢཎᅉ㑇ఏᏊࡀ༢㞳ࡉࢀ࡚࠸ࡿ㸬㑇ఏᙧ ᘧࡣᖖᰁⰍయඃᛶ㸦AD㸧ࡢࡶࡢࡀከࡃ㸪༙ᩘ௨ୖࡢ ࡛ཎᅉ㑇ఏᏊኚ␗ࡀྠᐃࡉࢀࡿࡀ㸪ᖖᰁⰍయຎᛶ 㸦AR㸧ࡢࡶࡢࡣᑡ࡞ࡃ㸪㒊ศࡣཎᅉ㑇ఏᏊࡸࡑࡢ ኚ␗ࡀ࡛᫂࠶ࡿ㸬 ᡃࠎࡣ㸪ᖖᰁⰍయຎᛶ㑇ఏ⪃࠼ࡽࢀࡿ㸪୧ぶࡀ ㏆ぶ፧࡛ྠ⬊Ⓨ⪅ࡀᏑᅾࡍࡿ௨ୗࡢᐙ⣔ࡘ࠸࡚㸪 ࡑࡢ㑇ఏᏊᗙཎᅉ㑇ఏᏊኚ␗ྠᐃࢆヨࡳ࡚࠸ࡿ㸬 㸺ᨭࡢෆᐜ㸼 ᖖᰁⰍయຎᛶ㑇ఏᛶ(AR)⪃࠼ࡽࢀࡿ㸪୧ぶࡀ㏆ ぶ፧࡛ྠ⬊Ⓨࡋ࡚࠸ࡿ㑇ఏᛶ②ᛶᑐ㯞⑷(HSP)ࡢ 2 ᐙ⣔ࡘ࠸࡚㸪ᮏᨭࡼࡾᐙ⣔ẖࡢ㐃㙐ゎᯒ㸪 ྛᐙ⣔ࡢᝈ⪅ 1 ྡࡘ࠸࡚࢚ࢡࢯ࣮࣒ゎᯒࢆ⾜ࡗ࡚ 㡬࠸ࡓ㸬ࡉࡽ㸪ࡢ AR ࡀࢃࢀࡿཎᅉ㑇ఏᏊᮍ ྠᐃࡢ HSP ᐙ⣔ೃ⿵㑇ఏᏊࡢኚ␗ࢆㄆࡵࡿ᳨ウ ࡋ࡚㡬࠸ࡓ㸬 activator-like effector nuclease (TALEN)ࢆ⏝࠸ࡓࢤࣀ ࣒⦅㞟ࢆ⾜࠸◚ቯࡋࡓࡀ㸪⾲⌧ᙧࡢኚࡣㄆࡵ࡞ ࡗࡓ㸬 ➨ 1 ࡢᐙ⣔ࡢࡶ࠺ 1 ྡࡢᝈ⪅ࡢ࢚ࢡࢯ࣮࣒ゎᯒࢆ ⾜ࡗࡓࡇࢁ㸪ᖖᰁⰍయඃᛶ㑇ఏᛶ(AD) HSP ࡛࠶ࡿ SPG3A ࡢ ཎ ᅉ 㑇 ఏ Ꮚ ATL1 ࡢ ᪤ ሗ ࡢ ⓗ ኚ ␗ (p.F413L)ࢆ࣊ࢸࣟ᥋ྜయ࡛ㄆࡵࡓ㸬ࡇࡢኚ␗ࡣᐙ⣔ ෆࡢูᝈ⪅ࡸᖖ⪅ࡣᏑᅾࡋ࡞ࡗࡓ㸬ࡲࡓ㸪ᐙ ⣔ෆࡢᝈ⪅ 2 ྡࡢࡳ ATL1 㑇ఏᏊࡢྠ⩏⨨ (p.Y459Y)ࢆ࣊ࢸࣟ᥋ྜయ࡛ㄆࡵࡓࡀ㸪ⓗព⩏ࡣ ࡛᫂࠶ࡿ㸬ᮏᐙ⣔ࡣぢࡅୖ㑇ఏᙧᘧࡣ AR ᛮࢃ ࢀࡓࡀ㸪AD-HSP ࡢ㑇ఏᏊኚ␗ࢆ᪂つ⏕ࡌࡓᝈ⪅ ࡀ ྡΰᅾࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡓ㸬 ➨ 2 ࡢᐙ⣔ࡣ㸪ᝈ⪅ 1 ྡᖖ⪅ 1 ྡ࡛ࡢ㐃㙐ゎ ᯒࡢ⤖ᯝ㸪➨ 1,2,3,6,8,10,11,14,15,19,22 ᰁⰍయ lod ್ࡢ㧗࠸㒊ศࢆㄆࡵࡓ㸦ᅗ 2㸧㸬 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ➨ 1 ࡢᐙ⣔ࡘ࠸࡚ࡣ㸪ᐙ⣔ෆ 5 ྡ㸦ᝈ⪅ 2 ྡ ᖖ⪅ 3 ྡ㸧ࡢ㐃㙐ゎᯒࡢ⤖ᯝ㸪➨ 5,6,13,17 ᰁⰍయ lod ್ࡢ㧗࠸㒊ศࢆㄆࡵࡓ㸦ᅗ 1㸧㸬 ࡑࡢᐙ⣔ࡢᝈ⪅ 1 ྡࡢ࢚ࢡࢯ࣮࣒ゎᯒࡼࡾ㸪ࡑ ࡢ㡿ᇦෆࡢ 2 ࡘࡢ㑇ఏᏊࡑࢀࡒࢀ࣍ࣔ᥋ྜయ࡛᪂ つࡢ୍ሷᇶ⨨ࢆྠᐃࡋࡓ㸬ࡑࡢ࠺ࡕ 1 ࡘࡢ㑇ఏᏊ F****ࡢኚ␗ࡣᐙ⣔ෆ࡛ඹศ㞳ࡀ☜ㄆࡉࢀࡓ㸬ࡑࢀࡣ ṇᖖࢥࣥࢺ࣮ࣟࣝ 208 ྡᏑᅾࡋ࡞ࡗࡓ㸬ࡉࡽ AR ࡀࢃࢀࡿ HSP ᐙ⣔ᝈ⪅ 89 ࡣ F****ࡢኚ␗ ࡣᏑᅾࡋ࡞ࡗࡓ㸬ࡇࢁࡀ㸪F****ࡢࢿࢵࢱࢶ࣓ ࢞ ࢚ ࣝ ࡢ ┦ ྠ 㑇 ఏ Ꮚ f**** ࢆ Transcription ࡑࡢᐙ⣔ࡢᝈ⪅ 1 ྡࡢ࢚ࢡࢯ࣮࣒ゎᯒࡼࡾ㸪ࡑ ࡢ㡿ᇦෆࡢ 6 ࡘࡢ㑇ఏᏊࡑࢀࡒࢀ࣍ࣔ᥋ྜయ࡛᪂ つࡢ୍ሷᇶ⨨ࢆྠᐃࡋࡓ㸬ࡑࡢෆ㸪ᐙ⣔ෆ࡛ඹศ 㞳ࡍࡿࡢࡣ㸪V*****㑇ఏᏊࡢኚ␗࡛࠶ࡗࡓ㸪ࡇࡢኚ ␗ࡣࢥࣥࢺ࣮ࣟࣝ 326 ྡࡣᏑᅾࡏࡎ㸪AR ࡀࢃࢀ ࡿ HSP ᐙ⣔ᝈ⪅ 89 13 ࡢ㏆ぶ፧ࡢ࠶ࡿ HSP ᐙ⣔ࡶ V*****ⓗኚ␗ࡣྠᐃࡉࢀ࡞ࡗࡓ㸬 V*****ࡢࢿࢵࢱࢶ࣓࢚࢞ࣝࡢ┦ྠ㑇ఏᏊ v***** ࢆ TALEN ࡛◚ቯࡋࡓࡇࢁ㸪࢜ࢱ࣐ࢪࣕࢡࢩࡢ㐠 ື␗ᖖࡀㄆࡵࡽࢀࡓ㸬 ࡉࡽ㸪➨ 2 ࡢᐙ⣔ࡢᐙ⣔ෆ 4 ྡ㸦ᝈ⪅ 2 ྡ ᖖ⪅ 2 ྡ㸧࡛ࡢ㐃㙐ゎᯒࡢ⤖ᯝ㸪➨ 1,2,10,11 ᰁⰍయ lod ್ࡀ㧗࠸㒊ศࡀ⤠ࡾ㎸ࡲࢀࡓ㸬ᐙ⣔ෆࡢࡶ࠺ 1 ྡࡢᝈ⪅ࡢ࢚ࢡࢯ࣮࣒ゎᯒ࡛ࡶ㸪᪤ሗࡢ㑇ఏᛶ②ᛶ ᑐ㯞⑷㑇ఏᏊࡢⓗኚ␗ࡣㄆࡵ࡞ࡗࡓ㸬 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᑠࡉ࡞ᑡᩘᐙ⣔ࢆ⏝࠸࡚ࡢཎᅉ㑇ఏᏊྠᐃᒎᮃ ࡀ㛤ࡅࡓ㸬➨ 1 ࡢᐙ⣔ࡣ SPG3A ᝈ⪅ࡀΰᅾࡋࡓᐙ⣔ - 374 - ࡛࠶ࡾ㸪F****ࡣཎᅉ㑇ఏᏊ࡛ࡣ࡞࠸ྍ⬟ᛶࡀ㧗࠸㸬 ➨ 2 ࡢᐙ⣔࡛ྠᐃࡉࢀࡓೃ⿵㑇ఏᏊ V*****ࡘ࠸ ࡚ࡣ㸪ᏙⓎࢆྵࡵࡓཎᅉ㑇ఏᏊᮍྠᐃࡢ᪂ࡓ࡞ᐙ ⣔㸪ᑐࡋ࡚ࡶ㑇ఏᏊኚ␗ࡢ᭷↓ࡢ᳨ウࡸ㸪ኚ ␗ࡼࡿ⣽⬊ෆᒁᅾࡢኚ࡞ࡶほᐹࡋ㸪┿ࡢཎᅉ 㑇ఏᏊ࡛࠶ࡿ᳨ドࢆ⾜ࡗ࡚ࡺࡃࡇࡋ࡚࠸ࡿ㸬 ⒪ἲ㛤Ⓨ㛵ࡍࡿ◊✲⌜㆟㸬 2014 ᖺ 1 ᭶ 8 ᪥㸪 ᮾி㸬 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᔱᓮᬕ㞝ࡽ㸬ᖖᰁⰍయຎᛶ㑇ఏᛶ②ᛶᑐ㯞⑷ࡢ᪂つ ཎᅉ㑇ఏᏊ᥈⣴㸬ᖹᡂ 25 ᖺᗘ㐠ືኻㄪࡢែゎ᫂ - 375 - බເᨭㄢ㢟㸸2010 ᖺᗘࠊ2011 ᖺᗘࠊ2013 ᖺᗘ ࢝ࢥࡑࡢ㏆⦕✀࠾ࡅࡿᐤ᳜≀㑅ᢥᶵᵓࡢ㐍 ᪂Ꮫ⾡㡿ᇦ◊✲ ᔱ⏣ ㏱࣭ᮾிᏛᏛ㝔㎰Ꮫ⏕⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢝ࢥࡣࢡ࣡ࡢⴥࡢࡳࢆ㣗㣵ࡍࡿࠋࢡ࣡ࡢⴥ ࡣ ࠊ 1-deoxynojirimycin 㸦 DNJ 㸧 ࡸ 1,4-dideoxy -1,4-imino-D-arabinitol㸦D-AB1㸧࡞ࡢ⢾㢮ఝࣝ ࢝ࣟࢻࡀ㔞ྵࡲࢀࠊ᳜㣗ᛶ᪻ᑐࡍࡿ㜵ᚚ ≀㉁ࡋ࡚ാ࠸࡚࠸ࡿࠋࡓ࠼ࡤࠊ࣐࣐ࣖࣘ࢞⛉ࡢ ࢚ࣜࢧࣥࡢ㣵 DNJ ࡸ D-AB1 ࢆῧຍࡍࡿࠊ⢾ࡢ ྾ࡸ௦ㅰࡀ㜼ᐖࡉࢀ࡚ᩘ᪥࡛Ṛஸࡍࡿࠋࡋࡋࠊ ࢝ࢥࡣ⢾㢮ఝࣝ࢝ࣟࢻࢆᦤྲྀࡋ࡚ࡶṇᖖᡂ 㛗ࡍࡿࡇࡀ࡛ࡁࡿࠋ⚾ࡓࡕࡣࠊ࢝ࢥࡀ⢾㢮ఝ ࣝ࢝ࣟࢻᢠᛶࡢᾘ㓝⣲ࢆᣢࡘࡇࡀࠊࢡ࣡ ࡢ㐺ᛂࡢࡓࡿᶵᵓ⪃࠼࡚࠾ࡾࠊᐇ㝿ࠊ㐍 ࡢ㐣⛬࡛⣽⳦ࡽ⋓ᚓࡋࡓᾘ㓝⣲șѸࣇࣝࢡࢺࣇ ࣛࣀࢩࢲ࣮ࢮ㸦ș-FFase㸧ࡀࣝ࢝ࣟࢻᢠᛶࡢ ࢫࢡ࣮ࣛࢮ࡛࠶ࡿࡇࢆ᫂ࡽࡋࡓ㸦Daimon et al., 2008, JBC㸧ࠋ ࢝ࢥ࡛ࡣ㸰ࡘࡢș-FFase 㑇ఏᏊࡢ࠺ࡕ BmSuc1 ࡀ୰⭠࡛ከ㔞Ⓨ⌧ࡋ࡚࠸ࡿࠋࡢ㫣⩟┠࡛᪻ࡶ ࡇࢀࡽࡢ࣮࢜ࢯࣟࢢࡀࢤࣀ࣒ୖᏑᅾࡍࡿࡀࠊࢡ࣡ 㣗࡛࠶ࡿ࢘ࢫࣂࢡ࣡ࢥࡸࢡ࣡ࣀ࣓࢞ࡢࡳ࡛㧗Ⓨ⌧ ࡋ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚ࠊ࢝ࢥࢆࡣࡌࡵࡍࡿࢡ࣡ 㣗᪻ࡣࠊ୰⭠࠾ࡅࡿ Suc1 ࡢⓎ⌧ࢆ㧗ࡵࡿࡇ࡛ࠊ ࢡ࣡ẘ⣲ࡢ㐺ᛂࢆᯝࡓࡋࡓ᥎ᐃ࡛ࡁࡿࠋࡲࡓࠊ ࢝ࢥ࢞⛉ࡣࢡ࣡⛉ࢡ࣡ᒓࢆ㣗ⲡࡍࡿ୍⩌㸦࢝ ࢥࠊࢡ࣡ࢥ➼㸧ࡢࠊ࢞ࢪ࣐ࣗࣝ࡞ࢳࢪࢡ ᒓࢆᐤ᳜≀ࡍࡿ୍⩌ࠊࡍ࡞ࢃࡕࢳࢪࢡ࢝ࢧࣥࠊ ࢸࣥ࢜ࣅࢩࣟ࢝ࢧࣥ➼ࡀᏑᅾࡍࡿࠋࡇࢀࡽࢡ࣡ࢆ ࠼ࡿᡂ㛗ࡀ㜼ᐖࡉࢀ࡚Ṛஸࡍࡿࠋࡇࢀࡽࢡ࣡㣗 ࡢ✀ࠊࢳࢪࢡᒓ㣗ࡢ✀ࢆẚ㍑ࡍࡿࡇࡼࡾࠊ ࢝ࢥ࢞⛉ࡢᐤࡢኚ㑄క࠺㐺ᛂ㐍ࡢ㐣⛬ࡀ᫂ ࡽ࡞ࡿࠋ ୍᪉ࠊ࢝ࢥࡣከࡃࡢ㣗ᛶኚ␗యࡀᏑᅾࡋࠊࡑ ࢀࡽࡣ᱓ⴥ௨እࡢ᳜≀࡛ࡶᦤ㣗ࡍࡿࠋ⚾ࡓࡕࡣࠊ࢝ ࢥࢤࣀ࣒ሗࢆά⏝ࡋࠊୡ⏺ඛ㥑ࡅ࡚㸰ࡘࡢኚ ␗య %W ࠾ࡼࡧ VSOL ࡢཎᅉ㑇ఏᏊೃ⿵ࢆྠᐃࡋࡓࠋ ࠸ࡎࢀࡶ 328࣓࣍࢜ࢻ࣓ࣥࢆᣢࡘ㌿ᅉᏊࢆࢥ࣮ ࢻࡋ࡚࠾ࡾࠊ㣗ᛶࡔࡅ࡛ࡣ࡞ࡃࠊࣇ࢙ࣟࣔࣥᛂ⟅ᛶ ࡶᨭ㓄ࡋ࡚࠸ࡓࠋࡋࡓࡀࡗ࡚ࠊࡇࡢศᏊࡀ᪂つ࡞ᐤ ᳜≀ࡢ㐍ฟ✀ศ࠶ࡓࡗ࡚ࠊ」ྜ㐺ᛂᙧ㉁ ࢆไᚚࡋ࡚࠸ࡿྍ⬟ᛶࡀ㧗࠸ࠋ࢝ࢥ㣗ᛶኚ␗యࡢ ཎᅉ㑇ఏᏊࡢゎ᫂ࠊ࠾ࡼࡧᶵ⬟ゎᯒࡽࠊ㣗ᛶ㐍 ࢆᨭ࠼ࡿ㑇ఏᏊࢿࢵࢺ࣮࣡ࢡࢆ᫂ࡽࡍࡿࠋ ୖグࡢ㸰ࡘࡢゅᗘࡽࠊ࢝ࢥࡢࢡ࣡㣗ࡢ㐺ᛂ 㐍㛵ࡍࡿ㑇ఏᏊ⩌ࡑࡢస⏝ᶵᵓࢆゎ᫂ࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 [ゎᯒ 1] ࢝ࢥࡑࡢ㏆⦕ࡢ᪻࠾ࡅࡿ㣗ᛶ㐍 㛵ࡍࡿჾᐁࡢẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ ࢝ࢥཬࡧ࢝ࢥ㏆⦕✀࡛࠶ࡿࢳࢪࢡ࢝ࢧࣥ Trilocha varians ࠊࢸࣥ࢜ࣅࢩࣟ࢝ࢧࣥ Ernolatia mooreiࠊ࢘ࢫࣂࢡ࣡ࢥ Rondotia mencianaࠊࢡ࣡ࢥ Bombyx mandarina 㸦௨ୖ࢝ࢥ࢞⛉㸧ࠊ࢚ࣜࢧࣥ Samia cynthia riciniࠊࢧࢡࢧࣥ Antheraea pernyi 㸦 ௨ ୖ ࣖ ࣐ ࣐ ࣘ ࢞ ⛉ 㸧ࠊ ࣚ ࢺ ࢘ ࢞ Mamestra brassicae 㸦 ࣖ ࢞ ⛉ 㸧ࠊ ࣓ ࣜ ࢝ ࢩ ࣟ ࣄ ࢺ ࣜ Hyphantria cunea 㸦ࣄࢺࣜ࢞⛉㸧ࠊࢡ࣡ࣀ࣓࢞ Glyphodes pyloalis㸦ࢶࢺ࢞⛉㸧ࡢྛ✀ࡽࠊࡑࢀࡒ ࢀ⤊㱋ᗂࡢ୰⭠࣭୰㒊⣒⭢࣭ᚋ㒊⣒⭢ࠊᡂࡢ㞤 ゐゅ࣭㞝ゐゅ࣭㞤ࣇ࢙ࣟࣔࣥ⭢ࡢ RNA ࢆㄪ〇ࡋࡓࠋ ࡇࢀ ࡽ RNA ࢆࠊ GA IIx ࠊ HiSeq2000 ࡲ ࡓࡣ HiSeq2500 ࡼࡿ RNA-seq ゎᯒ౪ࡋࡓࠋ [ゎᯒ 2] ࢝ࢥࡢ㣗ᛶኚ␗యࡢᗂᑠ㢞ࠊゐゅ࠾ࡼ ࡧ⬻ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࡢẚ㍑ ࢝ࢥṇᖖ⣔⤫㸦p50T㸧࠾ࡼࡧ㣗ᛶ␗ᖖ⣔⤫࠾ ࡅࡿᏛឤぬࡢ␗ᖖࡢཎᅉࢆ᥈ࡿࡓࡵࠊኚ␗⣔⤫ "spli"࠾ࡼࡧṇᖖ⣔⤫"p50T"ࡢࡑࢀࡒࢀࡘ࠸࡚ࠊ⤊ 㱋ᗂ㸦㞝㸧ࡢゐゅ࣭ᑠ㢞࠾ࡼࡧᡂ㸦㞤࣭㞝㸧ࡢ ゐゅ࣭⬻ࡽࠊRNA ࢆㄪ〇ࡋࡓࠋࡑࢀࡽ RNA ࢆࠊ GA IIX ࡼࡿ RNA-seq ゎᯒ౪ࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 >ゎᯒ@࢝ࢥࡑࡢ㏆⦕✀࠾ࡅࡿ㣗ᛶ㐍㛵 ࡍࡿჾᐁࡢẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ ᚓࡽࢀࡓ࢚࣌ࣥࢻࡢ࣮ࣜࢻࡣࠊ7ULQLW\ࡼࡾ ࢭࣥࣈࣝࡋࠊ56(0ࡼࡿ࣐ࢵࣆࣥࢢࢆ⾜ࡗࡓࠋ%/$67 ࡼࡾࢥࣥࢸࢢࡢࣀࢸ࣮ࢩࣙࣥࢆ⾜࠸࣮࢜ࢯࣟ ࢢ㛫ࡢⓎ⌧㔞ࢆ✀㛫࡛ẚ㍑ࡋࡓ⤖ᯝࠊࢡ࣡㣗᪻ࡢ ୰⭠࡛ࡣࠊ㠀ࢡ࣡㣗ࡢ᪻ẚ࡚ࠊșࣇࣝࢡࢺࣇ ࣛࣀࢩࢲ࣮ࢮ㑇ఏᏊ㸦6XF㸧ࡢⓎ⌧㔞ࡀ㧗ࡗࡓࠋ șࣇࣝࢡࢺࣇࣛࣀࢩࢲ࣮ࢮࡣࢡ࣡ࡢஙᾮẘ⣲࡛࠶ ࡿ⢾㢮ఝࣝ࢝ࣟࢻ⪏ᛶࡀ࠶ࡿࡇࡀ▱ࡽࢀ࡚ ࠸ࡿࠋࡉࡽࠊȘࢢࣝࢥࢩࢲ࣮ࢮᆺࡢྍ⁐ᛶࢫࢡࣛ ࣮ࢮࢆࢥ࣮ࢻࡍࡿ0DO࡞ࠊከᩘࡢ⢾ศゎ㓝⣲ࡢ Ⓨ⌧㔞ࡀࠊࢡ࣡㣗࡛᪻ቑຍࡋ࡚࠸ࡓࠋ᱓ⴥྵࡲ ࢀࡿ⢾㢮ఝࣝ࢝ࣟࢻࡣࠊȘࢢࣝࢥࢩࢲ࣮ࢮࢆ㜼 ᐖࡍࡿẘ⣲࡛࠶ࡿࡢ࡛ࠊࢡ࣡㣗᪻ࡢᾘ⟶࠾ࡅ ࡿ⢾ศゎ㓝⣲㑇ఏᏊࡢⓎ⌧㔞ୖ᪼ࡣࠊࢡ࣡ẘࡢ ᑐᢠᡭẁ࡛࠶ࡿ᥎ᐃࡉࢀࡿࠋ 51$VHTࢹ࣮ࢱࡽ᫂ࡽ࡞ࡗࡓせ࡞Șࢢࣝ ࢥࢩࢲ࣮ࢮࡢ0DO࠾ࡼࡧ6XKࡘ࠸࡚ࠊ⤌࠼ࢱࣥࣃ ࢡ㉁ࢆస〇ࡋࠊࡑࡢࢫࢡ࣮ࣛࢮάᛶ࠼ࡿ⢾㢮ఝ ࣝ࢝ࣟࢻࡢᙳ㡪ࢆㄪࡓࡇࢁࠊ࢝ࢥ࡞ࡢ ࢡ࣡㣗ࡢ᪻ࡢࢫࢡ࣮ࣛࢮࡣࠊ㠀ࢡ࣡㣗ࡢࢫࢡ࣮ࣛ ࢮࡼࡾࡶࣝ࢝ࣟࢻࡢᢠᛶࡀ㧗࠸ࡇࡀศ ࡗࡓࠋ >ゎᯒ@࢝ࢥࡢ㣗ᛶኚ␗యࡢᗂᑠ㢞ࠊゐゅ࠾ࡼ ࡧ⬻ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࡢẚ㍑ VSOLࡣࠊ᱓ⴥ௨እࡢ㣵ࢆᦤ㣗ࡋ࡚ࡋࡲ࠺㣗ᛶ␗ᖖ ࡔࡅ࡛࡞ࡃࠊᛶࣇ࢙࡛ࣟࣔࣥ࠶ࡿ࣎ࣥࣅࢥ࣮ࣝᑐ ࡍࡿᛂ⟅ᛶࡀపୗࡍࡿᙧ㉁ࢆ᭷ࡍࡿࠋ㞝ᡂゐゅࡢ - 376 - 51$VHTࢹ࣮ࢱࢆ࢝ࢥࢤࣀ࣒࣐ࢵࣆࣥࢢࡋࡓ⤖ ᯝࠊኚ␗య࡛ࡣ࣎ࣥࣅࢥ࣮ࣝཷᐜయ㑇ఏᏊ%P2UࡢⓎ ⌧ࡀ࢝ࢥṇᖖ⣔⤫㸦S7㸧ࡢᩘⓒศࡢపୗࡋ࡚ ࠸ࡿࡇࡀࢃࡗࡓࠋࡉࡽࠊࡑࡢࡢႥぬཷᐜయ 㑇ఏᏊࡢⓎ⌧ࡶࠊኚ␗య࡛ࡣῶᑡࡋ࡚࠸ࡓࠋVSOLࡣ 328࣓࣍࢜ࢻ࣓ࣥࢆࡶࡘ㌿ᅉᏊࡢ㑇ఏ㸦%PDFM㸧 ࡢḞኻኚ␗య࡛࠶ࡿࠋ㌿ᅉᏊ㸯ࡘࡢኚ࡛ᐤ㑅 ᢥ㓄അ⪅㑅ᢥࡢ୧᪉ࡀྠኚࡍࡿࡇࡽࠊ ✀ศࡢ㑇ఏᏊᶵᵓࢆㄝ᫂ྍ⬟࡞ࣔࢹࣝࡋ࡚ࠊ VSOLኚ␗యࡣ⯆῝࠸ᮦᩱ࡛࠶ࡿࡇࡀศࡗ࡚ࡁ ࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏゎᯒ࡛ᚓࡽࢀࡓྛ✀᪻ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮ ࣒ࡢẚ㍑ࡼࡗ࡚ࠊSuc1 㑇ఏᏊࢆࡣࡌࡵࡍࡿ⢾ ศゎ㓝⣲ࡢ㑇ఏᏊࡢⓎ⌧㔞ࡀࠊ㫣⩟┠᪻ࡢ࡞࡛ ࡶࠊ≉࢝ࢥ࠾ࡼࡧࢡ࣡㣗ࡢ✀≉␗ⓗከ࠸ࡇ ࢆぢ࠸ࡔࡋࡓࠋࢫࢡ࣮ࣛࢮࡸࢺࣞࣁ࣮ࣛࢮࡢࡼ࠺ ࡞⢾ศゎ㓝⣲ࡣࠊࢡ࣡ࡢஙᾮྵࡲࢀࡿẘ⣲ࡍ࡞ ࢃࡕ⢾㢮ఝࣝ࢝ࣟࢻࡢᶆⓗ࡛࠶ࡿࡇࡽࠊࢡ ࣡㣗ࡢ᪻ࡣࣝ࢝ࣟࢻࡢᑐᢠᡭẁࡋ࡚ࠊ ⢾ศゎ㓝⣲ࡢⓎ⌧㔞ࢆୖ᪼ࡉࡏ࡚࠸ࡿ⪃࠼ࡽࢀࡿࠋ ࡲࡓࠊ⤌࠼ࢱࣥࣃࢡ㉁ࢆస〇ࡍࡿᐇ㦂ࡢ⤖ᯝ ࡽࠊMal1 ࡸ Suh ࠸ࡗࡓȘ-ࢢࣝࢥࢩࢲ࣮ࢮᆺࡢࢫ ࢡ࣮ࣛࢮࡘ࠸࡚ࡣࠊࢡ࣡㣗ࡢ✀࡛≉␗ⓗ⢾㢮ఝ ࣝ࢝ࣟࢻᢠᛶࡀୖ᪼ࡋ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡘࡲࡾࠊࡇࢀࡽ㑇ఏᏊࡣࠊࢡ࣡ࡢ㐺ᛂ㐍 ࡢ㐣⛬࡛ࠊ㑇ఏᏊⓎ⌧ࡢୖ᪼࠸࠺㔞ⓗኚࡔࡅ ࡛࡞ࡃࠊᢠᛶ࠸࠺㉁ⓗ࡞ኚࡶ⋓ᚓࡋ࡚ࡁࡓࡶ ࡢ᥎ᐃࡉࢀࡿࠋ ᪂Ꮫ⾡㡿ᇦ◊✲ࠕ」ྜ㐺ᛂᙧ㉁㐍ࡢ㑇ఏᏊᇶ┙ ゎ࡛᫂ࠖࡣࠊ࢝ࢥ࢞⛉㏆⦕✀࡛࠶ࡾ࡞ࡀࡽ㠀ࢡ࣡ 㣗ࡢࢳࢪࢡ࢝ࢧࣥ࠾ࡼࡧ࢚ࣜࢧࣥࡢࢤࣀ࣒ゎㄞࢆ ᐇ⾜ࡋࡓࠋᮏ⏦ㄳ࡛ᚓࡽࢀࡓࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ ࢹ࣮ࢱࢆࠊࢤࣀ࣒ࡢ㓄ิ↷ྜࡍࡿࡇࡼࡾࠊ࢝ ࢥ࢞㏆⦕✀㸦࢝ࢥ࢞ୖ⛉㸧ࡢࢡ࣡㣗ࡢ㐺ᛂ ࠾࠸࡚ࠊ⢾ศゎ㓝⣲㑇ఏᏊࡸႥぬཷᐜయ㑇ఏᏊ࡞ ࡀ࠸ࡋ࡚㉁ⓗ࣭㔞ⓗኚࡋࡓࡢࠊศᏊ㐍 Ꮫⓗ࡞どⅬ࡛ㄪ࡚࠸ࡿࡇࢁ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 1. Wang, H., Kiuchi, T., Katsuma, S., and Shimada, T., A novel sucrose hydrolase from the bombycoid silkworms Bombyx mori, Trilocha varians, and Samia cynthia ricini with a substrate specificity for sucrose. Insect Biochemistry and Molecular Biology 61:46-52 ᮏㄢ㢟ࡣ௨ୗࡢ᪉ࠎࡢඹྠ◊✲࡛⾜ࢃࢀࡓ㸸㛫 㐍 1࣭ᮌෆ㝯ྐ 1࣭㛛㧗᫂ 1,2࣭⸨ ࿌ 1,3࣭⋤ ⳹ර 1,4࣭ᕝᮏ᐀Ꮥ 1࣭㛛⏣ᖾ 1࣭㕥ᮌ ✨ 5࣭Ⳣ㔝⣧ኵ 5 㸦1.ᮾ࣭㎰ࠊ2.⏕≀◊ࠊ3.⌧ࠊ4.⌧ύỤࠊ5. ᮾ࣭᪂㡿ᇦ㸧 - 377 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࢝ࢥ࠾ࡅࡿᛶỴᐃ࢝ࢫࢣ࣮ࢻ㑇ఏᏊ㔞⿵ṇ࠾ࡼࡧ⤌࠼ᢚไᶵᵓࡢࢡࣟࢫࢺ࣮ࢡ ᇶ┙◊✲㸦A㸧 ᔱ⏣ ㏱࣭ᮾிᏛᏛ㝔㎰Ꮫ⏕⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ື≀ࡢᛶỴᐃᵝᘧࡣࡁࢃࡵ࡚ከᵝ࡛࠶ࡾࠊ᪻ ࠾࠸࡚ࡶ✀ࡼࡗ࡚␗࡞ࡿᛶỴᐃᶵᵓࡀ᥇⏝ࡉࢀ࡚ ࠸ࡿࠋࢳࣙ࢘┠᪻ࡢᛶỴᐃࡣࠊྂࡃࡽ࢝ࢥ ࠾ࡅࡿ㑇ఏᏛⓗ◊✲ࢆ୰ᚰ㐍ࡵࡽࢀ࡚ࡁࡓࠋ࢝ ࢥࡣ W ᰁⰍయࡀᏑᅾࡍࢀࡤ㞤ࡍࡿࡇࡀ▱ࡽࢀ࡚ ࠸ࡓࡀࠊࡑࡢศᏊᐇయࡣ㛗ᖺᮍゎ᫂ࡢࡲࡲ࡛࠶ࡗࡓࠋ ࡋࡋ 2014 ᖺࠊᡃࠎࡢࢢ࣮ࣝࣉࡣࠊW ᰁⰍయୖࡢ ᑠศᏊ RNA ࡀ࢞ࢻ࡞ࡾࠊZ ᰁⰍయୖࡢ㞝㑇ఏ Ꮚ Masc ࡢ mRNA ࡀษ᩿ࡉࢀࡿࡇࡀࠊ࢝ࢥࡢᛶ Ỵᐃࡢ᭱ୖࡢ⤌ࡳ࡛࠶ࡿࡇࢆゎ᫂ࡋࡓ 㸦Kiuchi et al., Nature, 2014㸧ࠋࡉࡽࠊMasc ࢱࣥ ࣃࢡ㉁ࡀࠊ㞝ࡢࡳ࡞ࡽࡎࠊ㑇ఏᏊ㔞⿵ṇࢆࡶᨭ㓄 ࡍࡿࢱࣥࣃࢡ㉁࡛࠶ࡿࡇࡶⓎぢࡋࡓࠋᮏ◊✲࡛ࡣࠊ Masc ࢱࣥࣃࢡ㉁ࡀ㑇ఏᏊ㔞⿵ṇࢆᐇ⾜ࡍࡿศᏊ࣓ ࢝ࢽࢬ࣒ࢆゎ᫂ࡋࠊࢳࣙ࢘┠᪻࠾ࡅࡿ㑇ఏᏊ㔞 ⿵ṇࡢㇺࢆ᫂ࡽࡍࡿࠋ୍᪉ࠊ࢝ࢥࡀ㞤ࡢῶ ᩘศ࠾࠸࡚ᰁⰍయཫࢆ㉳ࡇࡉ࡞࠸࣓࢝ࢽࢬ࣒ ࢆㄪᰝࡍࡿࡇ࡛ࠊࢳࣙ࢘┠᪻࠾ࡅࡿᛶỴᐃ࢝ ࢫࢣ࣮ࢻ⤌࠼ࡢ㛵ಀࢆศᏊ࡛ࣞ࣋ࣝゎ᫂ࡍࡿࠋ ࡇࢀࡽࡢ┠ⓗࢆ㐩ᡂࡍࡿࡓࡵࠊMasc ࢱࣥࣃࢡ㉁ࡢ ᶆⓗ࡞ࡿ DNA ࡢሷᇶ㓄ิࢆࠊᇵ㣴⣽⬊࡛ Masc ࢆᙉไⓎ⌧ࡉࡏࠊChIP ゎᯒࢆ⾜࠺ࡇࡼࡗ࡚᫂ࡽ ࡋࡓ࠸ࠋࡲࡓࠊᛶỴᐃ࢝ࢫࢣ࣮ࢻ࡛ Masc ࡢୗ ὶ࠶ࡿ㑇ఏᏊ⩌ࢆࠊRNA-seq ࡼࡗ࡚᫂ࡽࡍ ࡿࠋࡉࡽࠊ㛫ᛶኚ␗య⣔⤫ࢆ⏝࠸࡚ࠊῶᩘศ ࠾ࡅࡿᛶ≉␗ⓗ࡞ཫࡢศᏊᶵᵓ㏕ࡿࠋ 㸺ᨭࡢෆᐜ㸼 (1) BmN4 ࢆࡗࡓ Masc ࡢ ChIP-seq 㸦9 ࢧࣥࣉࣝ㸧 ᮏ◊✲࡛ࡣࠊ㹘ᰁⰍయࢥ࣮ࢻࡉࢀࠊࢪࣥࢡࣇࣥ ࣮࢞ࢻ࣓ࣥࢆ᭷ࡍࡿࢱࣥࣃࢡ㉁ Masc ࡢᶵ⬟ࢆ᫂ ࡽࡍࡿࡇࡀࡁ࡞┠ᶆ࡛࠶ࡿࠋࡑࡢࡓࡵࠊ࢝ ࢥ༸ᕢ⏤᮶ࡢᇵ㣴⣽⬊࡛࠶ࡿ BmN ࠊ✀ࠎࡢᨵ ኚࢆࡇࡋࡓ Masc ࢆᙉไⓎ⌧ࡉࡏࠊࡑࢀࡀ⤖ྜ ࡍࡿࢤࣀ࣒㡿ᇦࡢሷᇶ㓄ิࢆࠊChIP ゎᯒࡼࡗ࡚᫂ ࡽࡍࡿࠋ (2) ๓㡯ࡢᐇ㦂ྠࡌࢧࣥࣉࣝࡢ RNA-seq㸦9 ࢧࣥ ࣉࣝ㸧 BmN ⣽⬊࠾ࡅࡿ Masc ࡢᙉไⓎ⌧ࡼࡿࢺࣛࣥ ࢫࢡࣜࣉࢺ࣮࣒ࡢኚࢆ᫂ࡽࡍࡿࡓࡵࠊ(1)ࡢྠ ࡌࢧࣥࣉࣝࡢ RNA ࢆ RNA-seq ࡼࡗ࡚ゎᯒࡍࡿࠋ (3) ṇᖖ⣔⤫㛫ᛶኚ␗⣔⤫ࡢ RNA-seq㸦8 ࢧࣥࣉ ࣝ㸧 㛫ᛶ⣔⤫ࡣࠊ㹕ᰁⰍయࡢኚ␗ࡼࡗ࡚ࠊ㞤ಶయࡢ ᙧ㉁ࡀ㒊ศⓗ㞝ࡋࡓ⣔⤫࡛࠶ࡿࠋᮏ◊✲࡛ࡣࠊ ṇᖖ⣔⤫㛫ᛶኚ␗⣔⤫ࡢࡑࢀࡒࢀࡘ࠸࡚ࠊ⏕Ṫ ᕢ㸦⢭ᕢ࣭༸ᕢ㸧ࡢ⤒ⓗ࡞ RNA-seq ゎᯒࢆ⾜࠸ࠊ ẚ㍑ࡍࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⌧ᅾࠊᨭࡼࡿゎᯒࢆᐇ୰ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ࡣࠊᡃࠎࡀ᫂ࡽࡋࡓ࢝ࢥ㹘ᰁⰍయୖ ࡢᛶỴᐃ㑇ఏᏊ Masc ࡢᶵ⬟ࢆࠊ㑇ఏᏊ࣭ศᏊࣞ࣋ ࡛ࣝලయⓗゎ᫂ࡍࡿࡇࢆࡵࡊࡋ࡚࠸ࡿࠋ ᨭෆᐜ(1)ࡣࠊMasc ࡢࢱ࣮ࢤࢵࢺ㑇ఏᏊࢆྠᐃ ࡋࠊ㞝࠾ࡼࡧ㑇ఏᏊ㔞⿵ṇࡢ࢝ࢫࢣ࣮ࢻࢆࠊศᏊ ࡛ࣞ࣋ࣝ᫂ࡽࡍࡿࡓࡵࡢࡶࡢ࡛࠶ࡿࠋMasc ࢱࣥ ࣃࢡ㉁ࡣ CCCH ࢱࣉࡢࢪࣥࢡࣇ࣮ࣥ࢞ࢻ࣓ࣥ ࢆ 2 ࡘᣢࡘ㞝ࢱࣥࣃࢡ㉁࡛࠶ࡾࠊ⬇Ꮚࢆ⏝࠸ࡓᐇ 㦂ࡽࠊ㑇ఏᏊ㔞⿵ṇ㛵ࢃࡿศᏊ࡛࠶ࡿࡇࡶุ ᫂ࡋ࡚࠸ࡿࠋᮏ◊✲࡛ࡣࠊᵝࠎ࡞ Masc ኚ␗య-GFP ⼥ྜ㑇ఏᏊࢆసᡂࡋࠊࡑࢀࡽࢆ BmN4 ⣽⬊ᑟධࡋ ࡚ ChIP-seq ࢆ⾜࠺ࡇ࡛ࠊMasc ࢱࣥࣃࢡ㉁ࡀ⤖ྜ ࡍࡿࢤࣀ࣒㡿ᇦࠊࡑࢀ㛵ࡍࡿ Masc ࡢᶵ⬟ࢻ ࣓ࣥࢆྠᐃࡍࡿࡇࡀ࡛ࡁࡿࠋࡑࡢ⤖ᯝࡽࠊ Masc ࢱࣥࣃࢡ㉁ࢆࡋࡓ㞝ࡸ㑇ఏᏊ㔞⿵ṇ࢝ࢫ ࢣ࣮ࢻࡢゎ᫂ࡀྍ⬟࡞ࡿࠋ ࡇࢀࡽࡼࡗ࡚ࠊࢩࣙ࢘ࢪࣙ࢘ࣂ࢚࡞ࡁࡃ ␗࡞ࡿᛶỴᐃᶵᵓࠊࡍ࡞ࢃࡕ piRNA ࢆ㉳Ⅼࡋࡓᛶ Ỵᐃ࢝ࢫࢣ࣮ࢻ㑇ఏᏊ㔞⿵ṇᶵᵓࡢㇺࡀゎ᫂ࡉ ࢀࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ ᮏㄢ㢟ࡣ௨ୗࡢ᪉ࠎࡢඹྠ◊✲࡛⾜ࢃࢀࡓ㸸㛫 㐍࣭ᮌෆ㝯ྐ࣭ᕝᮏ᐀Ꮥ㸦ᮾ࣭㎰㸧ࠊ㕥ᮌ ✨࣭Ⳣ 㔝⣧ኵ㸦ᮾ࣭᪂㡿ᇦ㸧 - 378 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࣞࢺࣟࣥ PV'1$ ࡼࡿཎᛶⓎ⌧ไᚚࢤࣀ࣒㐍࠾ࡅࡿᙺ ᇶ┙◊✲㸦㹁㸧 ᓥᮏ ᩚ࣭ᗈᓥᏛᏛ㝔⏕≀ᅪ⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⣽ ⳦ ࡢ ㏫ ㌿ 㓝 ⣲ 㸦 RT 㸧 ࡣ 㸪 multicopy single-stranded DNA 㸦 msDNA 㸧 ࡤ ࢀ ࡿ RNA-DNA 」 ྜ య ࡢ ྜ ᡂ ᚲ 㡲 ࡢ 㓝 ⣲ ࡛ ࠶ ࡿ ࠋ Vibrio cholerae ࡢ RT 㑇ఏᏊ㸦ret㸧ࡣࢤࣀ࣒ୖ࡛ msDNA ࢆࢥ࣮ࢻࡍࡿ㡿ᇦ㸦msr -msd㸧 2 ࡘࡢᶵ ⬟ᮍ▱ࡢ ORF㸦orf540㸪orf205㸧ࡶ㸪ࣞࢺࣟ ࣥࡤࢀࡿ࢜࣌ࣟࣥ㸦ࣞࢺࣟࣥ-Vc95㸧ࢆᙧᡂࡋ࡚ ࠸ࡿࠋࣞࢺࣟࣥ-Vc95 ࡢᶵ⬟ࡣゎ᫂ࡉࢀ࡚࠸࡞࠸ࡀ㸪 ὶ⾜ᛶࢥࣞࣛࡢཎᅉ࡞ࡿ V. cholerae O1/O139 ⾑ Ύᆺࡢࡍ࡚ࡢᰴࡀࣞࢺࣟࣥ-Vc95 ࢆಖ᭷ࡋ࡚࠸ࡿ ࡓࡵ㸪ཎᛶࣞࢺࣟࣥࡢ㛵ಀࡀᙉࡃ♧၀ࡉࢀ࡚࠸ ࡿࠋ ࡲࡓ㸪ࡇࢀࡲ࡛ࡢ◊✲࡛㸪ࣞࢺࣟࣥ-Vc95 ࡢ ret ࢆḞᦆࡉࡏࡓᰴ㸦TSC-11㸧㔝⏕ᰴ㸦MDO-6㸧࠾ࡼ ࡧḞᦆᰴࡢ┦⿵ᰴ㸦TSC-11/pBNR-1㸧ࡢ㑇ఏᏊⓎ ⌧ ࣞ࣋ࣝࡢ ẚ㍑ࡼࡾ 㸪 VC0176㸦 transcriptional regulator, putative 㸧 㑇 ఏ Ꮚ VC0177 㸦transcriptional regulator, VspR㸧㑇ఏᏊࡢⓎ⌧ࡀ ኚࡋ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ VC0176㸪 VC0177 ୧ 㑇 ఏ Ꮚ ࡣ VSP-1 㸦 Vibrio seventh pandemic island㸧ࡤࢀࡿ pathogenicity island ୖࢥ࣮ࢻࡉࢀ࡚࠸ࡿࡓࡵ㸪ࡇࡢ 2 ࡘࡢ㑇ఏᏊࡀࣞ ࢺࣟࣥࡲࡓࡣ msDNA ࢆࡋ࡚ཎᛶࢆㄪ⠇ࡋ࡚࠸ ࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋ VC0176㸪VC0177 ୧㑇ఏᏊࡢࣉ࣮ࣟࣔࢱ࣮㡿ᇦࡢ ሷᇶ㓄ิࢆㄪࡓࡇࢁ㸪msDNA ࡢ DNA ࢫࢸ࣒㒊 ศ㒊ศⓗ୍⮴ࡍࡿ㓄ิࡀᏑᅾࡋࡓࠋᙜ◊✲ᐊ࡛ ࡣ㸪msDNA ࡀࢹࢥ᰾㓟ࡢࡼ࠺ᶵ⬟ࡋ㸪㌿ไ ᚚᅉᏊ࡛࠶ࡿ VC0176 ࢱࣥࣃࢡ㉁⤖ྜࡍࡿࡇ ࡼࡗ࡚㸪VC0177 㑇ఏᏊࡢⓎ⌧ไᚚࢆ⾜ࡗ࡚࠸ࡿࡢ ࡛ࡣ࡞࠸࠸࠺௬ㄝࢆ❧࡚㸪ࢤࣝࢩࣇࢺἲࢆ⏝࠸ ᳨࡚ドࡋࡓࠋࡇࡢ᳨ド⤖ᯝࡼࡾ㸪VC0176 ࢱࣥࣃ ࢡ㉁ࡣ VC0177 㑇ఏᏊࡢࣉ࣮ࣟࣔࢱ࣮㡿ᇦ㸪࠾ࡼࡧ msDNA ࡢ DNA ࢫࢸ࣒㒊ศ⤖ྜࡍࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋ ୖ㏙ࡢࢤࣝࢩࣇࢺἲࡼࡿ⤖ᯝࡣ in vitro ࠾ࡅ ࡿ DNA VC0176 ࢱࣥࣃࢡ㉁ࡢ⤖ྜࡢ᭷↓ࢆ☜ㄆ ࡋࡓࡶࡢ࡛࠶ࡿࡀ㸪ᐇ㝿ࢥࣞࣛ⳦⣽⬊ෆ࡛ࡶྠᵝ ࡢ⤖ྜࡀ㉳ࡇࡾ㸪㑇ఏᏊࡢⓎ⌧ไᚚࢆ⾜ࡗ࡚࠸ࡿ ࠺ࡣ࡛᫂࠶ࡿࠋᅇ㸪in vivo ࠾࠸࡚ VC0176 ࢱࣥࣃࢡ㉁┦స⏝ࡍࡿ DNA ࡢ㒊ࢆ≉ᐃࡋ㸪 VC0176 ࡀไᚚࡍࡿ㑇ఏᏊ⩌ࢆ⥙⨶ⓗゎᯒࡍࡿࡇ ࡀᮏᨭ࡛ ChIP-Seq ゎᯒࢆ⾜࠺┤᥋ⓗ࡞┠ⓗ࡛ ࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 V. cholerae O1 El Tor P1418 ᰴ FLAG ࢱࢢࡢ ࠸ࡓ VC0176 ࢱࣥࣃࢡ㉁ࢆⓎ⌧ࡍࡿࣉࣛࢫ࣑ࢻࢆᑟ ධࡋ㸪ࣛࣅࣀ࣮ࢫ࡛ㄏᑟࡍࡿࡇࡼࡗ࡚ VC0176 ࢱࣥࣃࢡ㉁ࢆⓎ⌧ࡉࡏࡓࠋࡑࡋ࡚㸪ࢥࣥࢺ࣮ࣟࣝ ࡋ࡚࣋ࢡࢱ࣮㸦pBAD33㸧ࢆᑟධࡋࡓᰴࢆ⏝࠸㸪ࡑ ࢀࡒࢀㄏᑟࢆ⾜ࡗࡓࡶࡢㄏᑟࢆ⾜ࢃ࡞ࡗࡓࡶࡢ㸪 ィ㸲ࢧࣥࣉࣝࡘ࠸࡚ ChIP-seq ゎᯒࢆ⾜ࡗࡓࠋࡑࡋ ࡚㸪ᚓࡽࢀࡓ⤖ᯝࡘ࠸࡚ࢧࣥࣉࣝ㛫࡛ẚ㍑ࡍࡿࡇ ࡼࡗ࡚ VC0176 ࡀ⤖ྜࡍࡿᰁⰍయୖࡢ㡿ᇦࢆ᫂ ࡽࡋࡓࠋゎᯒࡣ㸪ᮾிᏛࡢ㕥ᮌ✨ᩍᤵⳢ㔝 ⣧ኵᩍᤵࡢ◊✲ᐊ౫㢗ࡋ㸪ᨭࢆ࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 VC0176 ࢱࣥࣃࢡ㉁ࡀ⤖ྜࡋࡓ㒊ࡽ 1 kb ௨ෆ ㌿㛤ጞ㒊㸦transcriptional start site, TSS㸧 ࡀᏑᅾࡍࡿ㑇ఏᏊࢆ᳨⣴ࡋࡓࡇࢁ㸪ࢥࣞࣛ⳦ࡢ 55 ࡢ㑇ఏᏊࡀヱᙜࡍࡿࡇࡀࢃࡗࡓࠋࡑࡢෆヂࡣ㸪 㓝⣲ࡢ㑇ఏᏊࡀ 33%㸪hypothetical protein ࡀ 27%㸪 ཎᛶ㛵㐃㑇ఏᏊࡀ 9%㸪ࢺࣛࣥࢫ࣏࣮ࢱ࣮ࡀ 4%㸪 ㌿ไᚚᅉᏊࡀ 2%㸪ࡑࡢࡀ 18%࡛࠶ࡗࡓࠋ ࡲ ࡓ 㸪 ヱ ᙜ 㑇 ఏ Ꮚ ࡢ ୰ DNA-directed RNA polymerase subunit ș㑇ఏᏊ㸦VC0328㸧ࡀྵࡲࢀ ࡚࠸ࡿࡇࡀࢃࡗࡓࠋูࡢᐇ㦂ࡽ msDNA ⤖ ྜࡍࡿࢱࣥࣃࢡ㉁ࡋ࡚ྠࡌࢱࣥࣃࢡ㉁ࡀྠᐃࡉࢀ ࡚࠸ࡿࠋࡇࡢࡇࡣ㸪msDNA ࠾ࡼࡧ VC0176 ࢱࣥ ࣃࢡ㉁ࡀ༠ྠࡋ࡚ RNA polymerase ࡢⓎ⌧㸪࠶ࡿ࠸ ࡣάᛶࡢㄪ⠇㛵ࡋ࡚࠸ࡿྍ⬟ᛶࢆ♧၀ࡋ࡚࠸ࡿࠋ ࡉࡽ㸪ࡑࢀࡼࡾࢥࣞࣛ⳦ࡢཎᛶࡢⓎ⌧ᐤ ࡋ࡚࠸ࡿࡢࡶࡋࢀ࡞࠸ࠋ᭦࡞ࡿゎᯒࡼࡾ㸪ࣞࢺ ࣟࣥࡸ msDNA ࡢᶵ⬟㸪࠾ࡼࡧࢥࣞࣛ⳦ࡢ᪂ࡓ࡞ ཎᛶⓎ⌧ㄪ⠇ᶵᵓࡢ୍➃ࢆ᫂ࡽࡍࡿࡇࡼࡾ㸪 ⛉◊㈝ㄢ㢟◊✲ࡢከ࡞㐍ᒎࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ChIP-Seq ゎᯒࡢ⤖ᯝ㸪msDNA ࡀ㛵ࡍࡿྍ⬟ᛶ ࡢ࠶ࡿ㌿ไᚚᅉᏊ VC0176 ࡼࡗ࡚ไᚚࡉࢀࡿೃ ⿵㑇ఏᏊࡋ࡚ᩘከࡃࡢ㑇ఏᏊࡀ᳨ฟࡉࢀࡓࠋᚋ㸪 ࡇࢀࡽࡢ㑇ఏᏊࡘ࠸࡚ಶࠎゎᯒࢆ㐍ࡵ㸪VC0176 ㌿ไᚚᅉᏊࡉࡽࡣ msDNA-Vc95 ࡢᶵ⬟ࡢゎᯒ ⤖ࡧࡘࡅࡿࡇࡀྍ⬟࡞ࡗࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 379 - බເᨭㄢ㢟㸸2011 ᖺᗘ ࢧࣥࢦඹ⏕ࡍࡿ〓⸴㢮ࡢẚ㍑ࢤࣀ࣒Ꮫⓗ◊✲ ᪂Ꮫ⾡㡿ᇦ◊✲ ᑘཱྀ ᰤ୍࣭Ἀ⦖⛉Ꮫᢏ⾡Ꮫ㝔Ꮫ࣐ࣜࣥࢤࣀ࣑ࢵࢡࢫࣘࢽࢵࢺ 㸺⫼ᬒ࣭┠ⓗ㸼 㠴ẟ⸴㢮ࡣࢧࣥࢦඹ⏕ࡍࡿ〓⸴ࠊ㉥₻ࡸ 㣗୰ẘࡢཎᅉ࡞ࡿ✀࡞⣙ 2000 ✀ࡀྵࡲࢀࠊᾏὒ ⎔ቃࢆ⪃࠼࡚࠸ࡃୖ࡛㔜せ࡞ࢢ࣮ࣝࣉ࡛࠶ࡿࠋࡲࡓ ᖖจ⦰ࡋࡓ᰾ࢆᣢࡘ࠸࠺ࡢ┿᰾⏕≀ࡣࡳࡽ ࢀ࡞࠸≉ᚩࢆഛ࠼ࠊࢤࣀ࣒ᵓ㐀ࡢ㐍ࢆ⪃࠼ࡿୖ࡛ ⯆῝࠸ࢢ࣮ࣝࣉ࡛ࡶ࠶ࡿࠋࡋࡋ࡞ࡀࡽࠊࡑࡢࢤ ࣀ࣒ࢧࢬࡣ 1.5 Gbp ࡽ 245 Gbp 㠀ᖖࡁ࠸ ࡇ࡞ࡽࠊࢤࣀ࣒ゎㄞࡣ࡞ࡉࢀ࡚࠸࡞ࡗࡓࠋ ࡑࡢࡼ࠺࡞⫼ᬒࡽࠊ 㠴ẟ⸴㢮࡛ึ࡞ࡾ࠺ࡿࠊ 〓⸴ࢤࣀ࣒ࢆỴᐃࡍࡿୖ࡛ࠊBAC ࡸ fosmid ࣛ ࣈ࣮ࣛࣜࡽࡢ㓄ิࢹ࣮ࢱࡀࠊ㧗⢭ᗘࡢࢤࣀ࣒ᴫせ 㓄ิࡢỴᐃᚲせ࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋࡉࡽࠊሗ ࿌ࡉࢀ࡚ࡁࡓ 㠴ẟ⸴ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࡸࣉ ࣮ࣟࣔࢱ࣮㡿ᇦࡢゎᯒࡽࠊࡑࡢ㑇ఏᏊⓎ⌧࣓࢝ࢽ ࢬ࣒ࡶ㠀ᖖ⊂≉࡞ࡶࡢ࡛࠶ࡿࡶࡋࢀ࡞࠸࠸࠺ ㆟ㄽࡀ࡞ࡉࢀ࡚࠸ࡓࠋࡑࢀࡺ࠼ࠊ㌿㛤ጞⅬ(TSS) ࢆண ࡍࡿࡓࡵࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒㓄ิࡢࢹ࣮ ࢱࡀࠊヲ⣽࡞㑇ఏᏊᵓ㐀ཬࡧⓎ⌧࣓࢝ࢽࢬ࣒ࡢゎᯒ 㠀ᖖ㔜せ࡞ࡶࡢ࡞ࡿ⪃࠼ࡽࢀࡓࠋ 㸺ᨭࡢෆᐜ㸼 1. 〓⸴ (Symbiodinium minutum)ࡢࢤࣀ࣒ DNA ࡽࠊBAC ࣛࣈ࣮ࣛࣜࢆసᡂࡋࠊ384 ࣉ࣮ࣞࢺࡢ 82 ᯛࣞࡋ࡚㡬࠸ࡓࠋࡑࡢ୰ࡢ 31,104 ࢡ࣮ࣟ ࣥࡢ࢚ࣥࢻࢩ࣮ࢣࣥࢫࢆࡋ࡚㡬࠸ࡓࠋ 2. 〓⸴ࡢࢤࣀ࣒ DNA ࡽࠊfosmid ࣛࣈ࣮ࣛࣜ ࢆసᡂࡋࠊ384 ࣉ࣮ࣞࢺࡢ 206 ᯛࣞࡋ࡚㡬࠸ ࡓࠋࡑࡢ୰ࡢ 19,200 ࢡ࣮ࣟࣥࡢ࢚ࣥࢻࢩ࣮ࢣࣥࢫࢆ ࡋ࡚㡬࠸ࡓࠋ 3. 〓⸴ࡢ mRNA ࡽࠊ㌿㛤ጞⅬ(TSS)ࢆண ࡍ ࡿࡓࡵࡢ RNAseq ࣛࣈ࣮ࣛࣜࢆసᡂࡋ࡚㡬࠸ࡓࠋ ࣑ࣝࢼࡢࢩ࣮ࢣࣥࢧ࣮ࡼࡾࠊࡑࡢࣛࣈ࣮ࣛࣜ ࡢࢩ࣮ࢣࣥࢫࢆ⾜ࡗ࡚㡬ࡁࠊ⣙ 29.1 Gbp ࡢ TSS 㓄 ิࢆỴᐃࡋ࡚㡬࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 1. ୡ⏺ඛ㥑ࡅ࡚ࠊ 㠴ẟ⸴㢮᰾ࢤࣀ࣒ࡢᴫせ㓄ิ ࢆỴᐃࡋࠊ㦫ࡁࡢ᰾ࢤࣀ࣒ᵓ㐀ࢆ᫂ࡽࡋࡓ (Shoguchi et al., 2013)ࠋBAC ࡸ fosmid ࡢ࢚࣌ࣥ ࢻ㓄ิࡣࠊࢭࣥࣈࣝࡋࡓࢤࣀ࣒ࡢṇ☜ᛶࢆ᳨ドࡍ ࡿୖ࡛㠀ᖖ᭷⏝࡛࠶ࡗࡓࠋTSS 㓄ิࢆࢤࣀ࣒ᴫせ 㓄ิ࣐ࢵࣆࣥࢢࡍࡿࡇࡼࡾࠊࢫࣉࣛࢫࢺࣜ ࣮ࢲ࣮ࢺࣛࣥࢫࢫࣉࣛࢩࣥࢢࢆཷࡅࡿࢺࣛࣥࢫࢡ ࣜࣉࢺ࣮࣒ࢆྠᐃࡍࡿࡇ࡛ࡁࡓࠋ 2. 〓⸴ࡢⴥ⥳యࢤࣀ࣒ࡣ࣑ࢽࢧ࣮ࢡࣝ DNA ࠸ ࠺≉Ṧ࡞ࢤࣀ࣒ᵓ㐀ࢆᣢࡘࠋTSS 㓄ิࢆⴥ⥳యࢤࣀ ࣒࣐ࢵࣆࣥࢢࡍࡿࡇࡼࡾࠊ TSS ୖὶᇦ࠾࠸ ࡚ࠊ12 ಶࡢ࣑ࢽࢧ࣮ࢡࣝ㛫࡛ಖᏑࡉࢀࡓ㠀ࢥ࣮ࢻ㡿 ᇦࢆྠᐃࡋࡓ(Mungpakdee et al., 2014)ࠋ 3. 㠴ẟ⸴㢮ࡢ࣑ࢺࢥࣥࢻࣜࢤࣀ࣒ࡣࠊᵝࠎ࡞㛗 ࡉࡢ᩿∦ࡀ⤌ࡳ࠼ࢆ㉳ࡇࡋࡓ≧ែ࡛Ꮡᅾࡋ࡚࠸ࡿ ࡓࡵࠊࡑࡢࢤࣀ࣒㓄ิࡣሗ࿌ࡉࢀ࡚࠸࡞ࡗࡓࠋ fosmid ࡢ࢚ࣥࢻ㓄ิ TSS 㓄ิࡢ࣐ࢵࣆࣥࢢࢹ࣮ ࢱࡶࡗ࡚ࠊ〓⸴࣑ࢺࢥࣥࢻࣜࡢࢤࣀ࣒㓄ิ Ỵᐃࡋࠊࡑࡢゎᯒ⤖ᯝࢆሗ࿌ࡋࡓ(Shoguchi et al., 2015)ࠋ 4. 㠴ẟ⸴ࡣࠊẘࢆྵࡴࣘࢽ࣮ࢡ࡞ḟ௦ㅰ⏘≀ 㸦࣏ࣜࢣࢳࢻ㸧ࢆྜᡂࡋࡑ࠺࡛࠶ࡿࡇࡀሗ࿌ࡉࢀ ࡚ࡁࡓࠋࡋࡋ࡞ࡀࡽࡑࡢ⏕ྜᡂࡣࠊ 㠴ẟ⸴ ඹ⏕ࡋ࡚࠸ࡿࣂࢡࢸࣜ࡞ࡀ㛵ࢃࡗ࡚࠸ࡿࡢࡶ ࡋ ࢀ࡞࠸ ࠸࠺ ၥ Ⅼࡀṧ ࡗ࡚࠸ ࡓࠋ〓 ⸴ S. minutum ࡢࢤࣀ࣒ࢥ࣮ࢻࡉࢀ࡚࠸ࡿ࣏ࣜࢣࢳࢳ ࢻྜᡂ㓝⣲ࡢ㑇ఏᏊ⩌ࢆゎᯒࡋࠊ〓⸴ࡀ࣏ࣜࢣࢳ ࢻࢆྜᡂࡋ࡚࠸ࡿࡇࢆࢧ࣏࣮ࢺࡍࡿ⤖ᯝࢆሗ࿌ࡋ ࡓ(Beedessee et al, 2015)ࠋTSS 㓄ิࡢ࣐ࢵࣆࣥࢢࢹ ࣮ࢱࡣࠊ࣏ࣜࢣࢳࢻྜᡂ㓝⣲㑇ఏᏊ㡿ᇦࡢヲ⣽࡞ゎ ᯒ᭷⏝࡛࠶ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 〓⸴ Symbiodinium ᒓࡣࠊࢧࣥࢦࢆྵࡴ่⬊ ື≀ࠊᾏ⥥ື≀ࠊ㌾యື≀ࠊᡥᙧື≀ࠊ᭷Ꮝඹ ⏕ࡍࡿ✀࡞ࡀྵࡲࢀࠊ㠀ᖖከᵝ࡛࠶ࡿࡇࡀሗ ࿌ࡉࢀ࡚ࡁ࡚࠸ࡿࠋSymbiodinium ࡢከᵝᛶࢆࢤࣀ ࣒ẚ㍑ࡢഃ㠃ࡽࡶࡳ࡚࠸ࡃࡇࡣࠊࢧࣥࢦࡢⓑ ⌧㇟ࢆྵࡴᾏὒ⎔ቃࢆ⪃࠼࡚࠸ࡃୖ࡛ࠊ㔜せ࡛࠶ࡿ ⪃࠼ࡽࢀࡿࠋᚓࡽࢀࡓࢤࣀ࣒㓄ิࢆཧ↷ࡍࡿࡇ ࡼࡾࠊࢧࣥࢦඹ⏕ࡋ࡚࠸ࡿ〓⸴ࡢࢤࣀ࣒ࡀࠊ ࢫࢺࣞࢫࡸ⎔ቃࡢኚᑐࡋࡢࡼ࠺ᛂࡋ࡚࠸ ࡿࡢࡀࠊẚ㍑ࢤࣀ࣒ࡸࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ ࡞ࡽ᫂ࡽ࡞ࡾࠊࢧࣥࢦࡢⓑ࣓࢝ࢽࢬ࣒ゎ ᫂ᑐ⟇❧㈉⊩ࡍࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 1. Shoguchi, E. et al., Curr. Biol. 23, 1399-1408 (2013) 2. Mungpakdee, S. et al., Genome Biol. Evol. 6, 1408-1422 (2014) 3. Shoguchi, E. et al., Genome Biol. Evol. 7, 2237-2244 (2015) 4. Beedessee, G. et al., BMC Genomics 16, 941 (2015) - 380 - බເᨭㄢ㢟㸸2013 ᖺᗘ ࢤࣀ࣒ࡢከᵝᛶゎᯒࡼࡿ⾲⌧ᆺྍረᛶࢆ᭷ࡍࡿ㔝⏕᳜≀㞟ᅋࡢ᱂ᇵ㛵ࡍࡿ◊✲ ⱝᡭ◊✲㸦㹀㸧 ⓑ⃝ ኴ࣭ࡎࡉ DNA ◊✲ᡤ ඛ➃◊✲㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ᱂ᇵ᳜≀ࡣ㔝⏕ࡢ᳜≀✀ẚ࡚⏕㛗㔞⦾Ṫຊ ࡀຎࡿࡇࡀከ࠸ࡀࠊࡇࢀࡣ᳜≀ࡀ᱂ᇵࡢ㐣⛬࡛ 㑇ఏⓗ࡞ከᵝᛶࠊࡑࢀక࠺ᵝࠎ࡞⎔ቃࢫࢺࣞࢫ ᑐࡍࡿᢠᛶࠊࡍ࡞ࢃࡕྍረᛶࢆኻࡗࡓࡇ㉳ ᅉࡍࡿ⪃࠼ࡽࢀ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚㏫ㄝⓗࠊ㑇 ఏⓗ࡞ከᵝᛶࢆ⥔ᣢࡋࡓࡲࡲ㔝⏕᳜≀ࢆ᱂ᇵ࡛ࡁ ࢀࡤࠊ㧗ᗘ࡞ྍረᛶࢆᣢࡗࡓ᱂ᇵ᳜≀ࡀసฟ࡛ࡁࡿ ྍ⬟ᛶࡀ㧗࠸ࠋࡘࡲࡾࠊࡇࢀࡲ࡛ே㢮ࡀᩘ༓ᖺࡶ ࡅ࡚⾜࡞ࡗ࡚ࡁࡓ㔝⏕᳜≀ࡢ᱂ᇵࢆࠊ⌧௦࠾࠸ ࡚ࡶࢤࣀ࣒ゎᯒᢏ⾡ࢆ㥑ࡍࡿࡇ࡛▷ᮇ㛫ࠊ ࡘࠊࡼࡾຠᯝⓗ㐩ᡂࡍࡿࡇࡀྍ⬟࡞ࡿࠋ 㔝⏕᳜≀㞟ᅋࡢࢤࣀ࣒ࡢከᵝᛶࢆඛ➃ⓗ࡞ࢤࣀ࣒ ゎᯒᢏ⾡ࡼࡾ᫂ࡽࡋࠊ᱂ᇵ㛵㐃㑇ఏᏊᗙࢆ ᅛᐃࡋࡓ㞟ᅋࡢ⾲⌧ᆺྍረᛶࢆホ౯ࡍࡿࡇࡀ࡛ ࡁࢀࡤࠊ᳜≀㞟ᅋࡢ㑇ఏⓗከᵝᛶࠊ⾲⌧ᆺࡢᅛᐃᗘࠊ ࠾ࡼࡧྍረᛶࡢ㛵㐃ࢆゎ࡛᫂ࡁࡿࡼ࠺࡞ࡿࠋ ࡢࢩ࣮ࢡ࢚ࣥࢫࣛࣈࣛࣜࢆస〇ࡋࡓࠋ࢚࣌ࣥࢻ ࣓ࢺ࣌ࣛࣈࣛࣜࡢ㓄ิศᯒࡣ Illumina ♫ࡢ HiSeq ࢆࠊࣟࣥࢢ࣮ࣜࢻࣛࣈࣛࣜࡢ㓄ิศᯒࡣ PacBio ♫ࡢ RS ࢆ⏝ࡋࡓࠋࡇࡢ࠺ࡕࠊ2 ✀㢮ࡢ࣓ ࢺ࣌ࣛࣈࣛࣜࣟࣥࢢ࣮ࣜࢻࣛࣈࣛࣜࡢస 〇ࠊཬࡧ㓄ิศᯒࡣࠕࢤࣀ࣒ᨭࠖࡢࢧ࣏࣮ࢺࢆཷ ࡅ࡚ᐇࡋࡓࠋᚓࡽࢀࡓ㓄ิࢹ࣮ࢱࢆရ㉁࡛⢭ᰝᚋࠊ ࡇࢀࢆࢭࣥࣈࣝࡍࡿࡇ࡛᥎ᐃࢤࣀ࣒㛗ࡢ 80%௨ ୖࢆ࢝ࣂ࣮ࡍࡿᴫせ㓄ิࢆᚓࡓࠋࡇࡢ㓄ิࡣࠊ┿᰾ ⏕≀ඹ㏻ࡢ㑇ఏᏊࢭࢵࢺࡢ 90%௨ୖࢆྵࢇࡔࡇ ࡽࠊࢤࣀ࣒ࡢ࢝ࣂ࣮⋡ࡣ༑ศ࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋ ࣀࢸ࣮ࢩࣙࣥ ᵓ⠏ࡋࡓࢤࣀ࣒㓄ิྵࡲࢀࡿࢺࣛࣥࢫ࣏ࢰࣥ➼ ࡢࣜࣆ࣮ࢺ㓄ิࡢ᳨ฟࢆ⾜࠸ࠊࣖࣁࢬ࢚ࣥࢻ࢘ࡢࢤ ࣀ࣒㓄ิࡢ 80%௨ୖࡀࣜࣆ࣮ࢺ㓄ิ࡛༨ࡵࡽࢀ࡚࠸ ࡿࡇࢆ᫂ࡽࡋࡓࠋ ࡉࡽࠊᵓ⠏ࡋࡓࢤࣀ࣒㓄ิࡽ㑇ఏᏊࢆண ࡍ ࡿࡓࡵࠊᶆ‽ᰴࡽⴥࠊⱼࠊ࠾ࡼࡧ᰿࡞ࡢྜィ ჾᐁࢆ᥇ྲྀࡋࠊࡇࢀࡽࡽ 51$ ࢆᢳฟࡋࠊࢩ࣮ ࢡ࢚ࣥࢫࣛࣈࣛࣜࢆసᡂࡋࠊIllumina ♫ࡢ MiSeq ࢆ⏝ࡋ࡚㓄ิศᯒࢆ⾜ࡗࡓࠋᚓࡽࢀࡓሗࠊ㏆ ⦕࣐࣓⛉᳜≀ࡢ㑇ఏᏊ㓄ิ➼ࢆ⏝ࡋ࡚㑇ఏᏊண ࢆ⾜࠸ࠊᵓ⠏ࡋࡓࢤࣀ࣒㓄ิୖ ௨ୖࡢ㑇ఏᏊ ࢆぢฟࡋࡓࠋࡇࢀࡽࡢ㑇ఏᏊ㓄ิࡣࠊ㏆⦕࣐࣓⛉᳜ ≀ࡢ㑇ఏᏊ㓄ิẚ㍑ࡋࠊࣖࣁࢬ࢚ࣥࢻ࢘≉᭷ࡢ ⱝᡭ◊✲㸦㹀㸧࡛ࡣࠊ㑇ఏⓗከᵝᛶࡢ㧗ࡉࢆ⥔ ᣢࡍࡿ㔝⏕᳜≀㞟ᅋ᱂ᇵ≉ᛶࢆࡍࡿࡇ࡛ࠊ 㧗ᗘ࡞ྍረᛶࢆᣢࡗࡓ᱂ᇵ᳜≀ࠊࡍ࡞ࢃࡕྛ✀ࡢ ࢫࢺࣞࢫ⪏ᛶࢆᣢࡗࡓ᱂ᇵ᳜≀ࡀ㛤Ⓨ࡛ࡁࡿྍ ⬟ᛶࢆ᳨ドࡍࡿࡇࢆ┠ⓗࡋ࡚࠸ࡿࠋࡑࡢࡓࡵ ࡣࠊ㔝⏕᳜≀᱂ᇵ≉ᛶࢆࡉࡏࡿࡓࡵᚲ せ࡞ࢤࣀ࣒㡿ᇦࠊ᱂ᇵ᳜≀㧗ᗘ࡞ྍረᛶࢆᣢ ࡓࡏࡿࡓࡵᚲせ࡞ࢤࣀ࣒㡿ᇦࡢ᪉ࡢሗྲྀᚓ ࡀྍḞ࡛࠶ࡿࠋࡑࡇ࡛ᮏᨭ࡛ࡣࠊゎᯒࡢᑐ㇟ ࡍࡿࣖࣁࢬ࢚ࣥࢻ࢘ࡢࢤࣀ࣒ᴫせ㓄ิࡢỴᐃࠊ 㑇ఏᏊࢆྠᐃࡋࡓࠋ 㔝⏕㞟ᅋࡢ㑇ఏⓗከᵝᛶࢆ⾲ࡋࡓࣁࢵࣉ࣐ࢵࣉࡢ సᡂࢆ┠ⓗࡋࡓࠋ 㔝⏕㞟ᅋࡢከᵝᛶゎᯒ 㸺ᨭࡢෆᐜ㸼 (1) 1 ಶయࡢࣖࣁࢬ࢚ࣥࢻ࢘ࡢࢩࣙࢵࢺ࢞ࣥࣛࣈ ࣮ࣛࣜస〇 PacBio RS ࡢ CLR ࣮ࣔࢻ࡛ࡢ㓄ิ ศᯒ (2) 1 ಶయࡢࣖࣁࢬ࢚ࣥࢻ࢘ࡢ࣓ࢺ࣌ࣛࣈࣛ ࣜࡢస〇㓄ิศᯒ (3) 768 ಶయࡢࣖࣁࢬ࢚ࣥࢻ࢘ࡢ࢚࣌ࣥࢻࣛࣈ ࣛࣜస〇㓄ิศᯒ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣖࣁࢬ࢚ࣥࢻ࢘ࢤࣀ࣒ࡢ㓄ิࢹ࣮ࢱࡢ㞟ࢭࣥ ࣈࣝ 㔝⏕᳜≀᱂ᇵ≉ᛶࢆࡉࡏࡿࡓࡵᚲせ࡞ࢤ ࣀ࣒㡿ᇦࠊ᱂ᇵ᳜≀㧗ᗘ࡞ྍረᛶࢆᣢࡓࡏࡿࡓ ࡵᚲせ࡞ࢤࣀ࣒㡿ᇦࡢ᪉ࡢሗࢆᚓࡿࡓࡵࡣࠊ ࣖࣁࢬ࢚ࣥࢻ࢘ࡢࢤࣀ࣒ゎㄞࡀᚲせ࡛࠶ࡿࠋࡑࡇ࡛ࠊ ࣖࣁࢬ࢚ࣥࢻ࢘ࡢ⣧⣔ᶆ‽ᰴࢆ☜❧ࡋࡓࠋ ᶆ‽ᰴࡽࢤࣀ࣒ DNA ࢆᢳฟࡋࠊ࢚࣌ࣥࢻ 㸦ࣥࢧ࣮ࢺ㛗 500 bp㸧ࠊ࣓ࢺ࣌㸦ࣥࢧ࣮ࢺ㛗 2 kbࠊ5 kbࠊ10 kbࠊཬࡧ 15 kb㸧ࠊཬࡧࣟࣥࢢ࣮ࣜࢻ ᙜึࡣࠊᅜྛᆅࡽ㞟ࡋࡓ 8 㞟ᅋࡢࣖࣁࢬ࢚ ࣥࢻ࢘ࡢ࢚࣌ࣥࢻࣛࣈࣛࣜࡢస〇㓄ิศᯒࢆ ィ⏬ࡋ࡚࠸ࡓࡀࠊࡑࡢᚋࡢࢩ࣮ࢡ࢚ࣥࢫᢏ⾡ࡢ㐍Ṍ ศᯒࢥࢫࢺࡢపୗక࠸ࠊゎᯒࡢᑐ㇟ࡍࡿ㞟ᅋ ᩘࢆ 12 㞟ᅋࡋࡓࠋྛ㞟ᅋ࣭ಶయࢆ RAD-Seq ἲ࡛ ศᯒࡋࠊᚓࡽࢀࡓ㓄ิࢹ࣮ࢱࢆᵓ⠏ࡋࡓࢤࣀ࣒㓄ิ ẚ㍑ࡍࡿࡇ࡛ࠊ㞟ᅋ࠶ࡓࡾ 5 ༓ࠥ1 ⛬ᗘࡢ SNP ࢆ᳨ฟࡋࠊ㔝⏕㞟ᅋࡢ㑇ఏⓗከᵝᛶࢆ⾲ࡋࡓࣁࢵࣉ ࣐ࢵࣉࢆస〇ࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᨭࡼࡗ࡚ࠊ㔝⏕㞟ᅋᬑ㐢ⓗྍረᛶࢆࡶࡓ ࡽࡍࢤࣀ࣒㡿ᇦࢆ㧗⢭ᗘ᥎ᐃࡍࡿࡓࡵࡢ㐨➽ࡀᚓ ࡽࢀࡓࠋḟⓗࠊࣁࢵࣉ࣐ࢵࣉࡽᚓࡽࢀࡿ㐃㙐 ᖹ⾮ࡢሗࢆ⏝ࡋࡓࢤࣀ࣒࣐ࢵࣆࣥࢢࡢ᪉ἲࡢ ☜❧ࡀぢ㎸ࡲࢀࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ‽ഛ୰ࠋ - 381 - බເᨭㄢ㢟㸸2012 ᖺᗘ ࢡ࣐ࣟࢳ࣮ࣥࣝࣉᵓ㐀ኚࡼࡿ⤌⧊≉␗ⓗ Shh Ⓨ⌧ไᚚࢩࢫࢸ࣒ ᇶ┙◊✲㸦㸿㸧 ᇛ▼ ಇᙪ࣭ሗ࣭ࢩࢫࢸ࣒◊✲ᶵᵓᅜ❧㑇ఏᏛ◊✲ᡤ ⣔⤫⏕≀◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 㑇ఏᏊࡢ⤌⧊≉␗ⓗࠊᮇ≉␗ⓗⓎ⌧ࡣࠊᰁⰍ యࡢ୕ḟඖᵓ㐀ࡢኚࡀࡁࡃ㛵ࢃࡗ࡚࠸ࡿࠋᰁⰍ యࡢ࣮ࣝࣉᵓ㐀ࡣࠊ࢚ࣥࣁࣥࢧ࣮࣭ࣉ࣮ࣟࣔࢱ࣮ࡢ ┦స⏝ࢆಁ㐍ࡍࡿࡇ࡛㌿ࡢάᛶ࣏ࢪࢸ ࣈ࡞ຠᯝࢆࡶࡓࡽࡍ㠃ࠊ≀⌮ⓗ࡞㞀ቨ࡞ࡾࠊ㌿ ࢿ࢞ࢸࣈ࡞ᙳ㡪ࢆ࠼ࡿࡇࡶ࠶ࡿࠋࡋࡋࠊ ᰁⰍయୖ୪ࢇࡔ㑇ఏᏊࡢ㛫࡛࣮ࣝࣉࡢᵓ㐀ኚࡀ ࠸ࡗࡓ࠸ࡢࡼ࠺⤌⧊ࡉࢀ࡚࠸ࡿࡢࠊࡑࡢ࣓ ࢝ࢽࢬ࣒ࡣ࠶ࡲࡾࡼࡃศࡗ࡚࠸࡞࠸ࠋ Shh 㑇ఏᏊᗙ࡛ࡣࠊ⣙ 1 Mb ཬࡪᗈ࡞ࢤࣀ࣒ 㡿ᇦ 10 ௨ୖࡢ࢚ࣥࣁࣥࢧ࣮ࡀࡍ࡛ྠᐃࡉࢀ࡚ ࠾ࡾࠊ⤌⧊≉␗ⓗ࡞Ⓨ⌧ไᚚᶵᵓࢆゎᯒࡍࡿࡓࡵࡢ Ⰻ࠸ࣔࢹࣝ㡿ᇦ࡞ࡗ࡚࠸ࡿࠋᡃࠎࡣࠊࡇࢀࡽࡢ࢚ ࣥࣁࣥࢧ࣮ࡀᩘⓒ Kb ཬࡪ㛗࠸㊥㞳ࢆ㉸࠼࡚ࠊ⤌⧊ ≉␗ⓗ Shh ࣉ࣮ࣟࣔࢱ࣮స⏝ࡍࡿࡇࢆࡍ࡛ ᫂ࡽࡋ࡚࠸ࡿࠋࡓࡔࡋࠊ࢚ࣥࣁࣥࢧ࣮ࢆḞኻࡋ ࡓ࣐࢘ࢫࡢ⫥ⱆ࡛ࡶ㔝⏕ᆺྠᵝ࡞ᰁⰍయືែࡀほ ᐹࡉࢀࡿࡇࡽࠊᰁⰍయᵓ㐀ࡢኚࡣࠊ࢚ࣥࣁ ࣥࢧ࣮ࡣูᮍ▱ࡢࢩࢫ㓄ิࢺࣛࣥࢫᅉᏊࡀᚲ せࡔ᥎ ࡉࢀࡿࠋ 㸺ᨭࡢෆᐜ㸼 (1) ChIP-seq 㑇ఏᏊⓎ⌧ࡢάᛶ┦㛵ࡋ࡚≉ᐃࡢࢤࣀ࣒㡿ᇦ ࢆࡘ࡞ࡄࡇࡀ▱ࡽࢀ࡚࠸ࡿ CTCF Cohesin 㛵 ࡋ ࡚ ࠊ Shh 㑇 ఏ Ꮚ ᗙ ୖ ࡢ ศ ᕸ ࢆ ▱ ࡿ ࡓ ࡵ ChIP-Seq ゎᯒࢆ⾜ࡗࡓࠋ࣐࢘ࢫࡢ 11.5 ᪥⬇ࡼࡾࠊ ⫥ⱆ㢌㒊ࡢ 2 ⤌⧊ࢆࡑࢀࡒࢀㄪ〇ࡋࠊCTCFࠊ Cohesin ࢧࣈࣘࢽࢵࢺࡢ Smc1aࠊCohesin ࡢ࣮ࣟࢹ ࣥࢢᅉᏊ࡛࠶ࡿ Mau2 ᑐࡍࡿ 3 ✀㢮ࡢ≉␗ᢠయ ࢆ⏝࠸࡚ࢡ࣐ࣟࢳࣥචỿ㝆ࢆ⾜ࡗࡓࠋ2 ⤌⧊ࡢච ỿ㝆 DNA ࣥࣉࢵࢺ DNAࠊィ 8 ࢧࣥࣉࣝ㛵ࡋ ࡚ HiSeq ࡼࡿ⥙⨶ⓗ㓄ิゎᯒࡢᨭࢆ࠾㢪࠸ࡋࡓࠋ (2) 4C-seq Chromosome Conformation Capture (3C)ἲࢆᛂ ⏝ࡋࡓ 4C ἲࡼࡾࠊ࣐࢘ࢫ⬇ Shh 㑇ఏᏊᗙ࿘㎶ࡢ ᰁⰍయ┦స⏝ࢆホ౯ࡋࡓࠋ࣐࢘ࢫ 11.5 ᪥⬇ࡢ Shh Ⓨ⌧⫥ⱆ⣽⬊ࡽࠊᰁⰍయ┦స⏝㒊ࢆྵࡴ⎔≧ ࡢ࣓࢟ࣛ DNA ࢆㄪ〇ࡋࠊྛ࣋ࢺ㓄ิタィࡋࡓ ࣉ࣐࣮ࣛࡼࡿࣥࣂ࣮ࢫ PCR ࢆ⾜ࡗࡓࠋ࣋ࢺ 㓄ิࡋ࡚⏝࠸ࡓࡢࡣࠊ௨ୗࡢ Shh 㑇ఏᏊᗙ࿘㎶ࡢ 7 ࣨᡤ࡛࠶ࡿࠋձShh ࣉ࣮ࣟࣔࢱ࣮㡿ᇦࠊղ⫥ⱆ࢚ ࣥࣁࣥࢧ࣮㡿ᇦࠊճ⫥ⱆ࢚ࣥࣁࣥࢧ࣮㏆ഐࡢ㐍ⓗ ಖᏑ㓄ิࠊ մShh ⩻ヂ㓄ิୖὶࡢ CTCF ⤖ྜࢧࢺࠊ յShh ⩻ヂ㓄ิୗὶࡢ CTCF ⤖ྜࢧࢺࠊնࣄࢺḞ ⫥㸦Achieropodia㸧ࡢཎᅉೃ⿵㡿ᇦࠊշShh ㏆ഐ ⨨ࡍࡿ Lmbr1 㑇ఏᏊࡢࣉ࣮ࣟࣔࢱ࣮ࠋࡇࡢ 7 ✀㢮ࡢ PCR ⏘≀ᑐࡋ࡚ࠊHiSeq ࡼࡿ⥙⨶ⓗ㓄ิ ゎᯒࢆ࠾㢪࠸ࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ChIP-Seq ἲࡼࡾࠊ࣐࢘ࢫ⬇⤌⧊࠾ࡅࡿ CTCF Cohesin 㛵㐃ࢱࣥࣃࢡ㉁ࡢࢤࣀ࣒ୖࡢศᕸࢆ⥙ ⨶ⓗ࣐ࢵࣉࡍࡿࡇࡀ࡛ࡁࡓࠋ⫥ⱆ࡛ࡢ Shh Ⓨ⌧ ᚲ㡲࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ CTCF ࡣࠊShh ⩻ ヂ㡿ᇦࢆᣳࡴࡼ࠺ୖὶ 3.5 Kb ୗὶ 6.5 Kb ࡢ ⨨ࣆ࣮ࢡࡀㄆࡵࡽࢀࡓࠋࡲࡓࠊ⫥ⱆ࢚ࣥࣁࣥࢧ࣮ ࡢୖὶୗὶ 20 Kb ྛࠎࣆ࣮ࢡࡀㄆࡵࡽࢀࡓࠋ୍ ᪉࡛ࠊ࢚ࣥࣁࣥࢧ࣮ Shh ⩻ヂ㡿ᇦࡢ㛫ࡢ 800 Kb ཬࡪࣥࢱ࣮ࢪ࣮ࣥ㡿ᇦ࡛ࡣࠊᩘಶࡢప࠸ࣆ࣮ࢡ ࡀㄆࡵࡽࢀࡿࡔࡅ࡛࠶ࡾࠊCTCF ࡢ⤖ྜࡣ MFCS1࣭ Shh ⩻ヂ㡿ᇦ≉␗ⓗ࡛࠶ࡗࡓࠋCTCF ࡣ࣍ࣔ㔞య ࢆᙧᡂࡍࡿࡇ࡛ࠊᰁⰍయ࣮ࣝࣉᵓ㐀ࡢᙧᡂᐤ ࡍࡿࡓࡵࠊChIP-seq ࡢ⤖ᯝࡣ⫥ⱆ࡛ࡢ Shh ࡢ㌿ άᛶ CTCF ࡀ㛵ࡍࡿྍ⬟ᛶࢆᙉࡃ♧၀ࡋ࡚࠸ ࡿࠋ ࡲࡓࠊ4C-Seq ἲࡼࡿゎᯒࡢ⤖ᯝࠊShh ⩻ヂ㡿ᇦ MFCS1 ࢆྵࡴ 1 Mb ㉸ࡢ༊⏬ࡢ୰࡛㧗㢖ᗘ┦ స⏝ࡀ㉳ࡇࡗ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࢀ ࡣࠊShh MFCS1 ࡀ≉ᐃࡢᰁⰍయ࣮ࣝࣉᵓ㐀ࢆస ࡗ࡚Ᏻᐃ࡞⤖ྜࢆࡋ࡚࠸ࡿ࠸࠺ࡼࡾࡶᰁⰍయୖࡢ ᶵ⬟ⓗ࡞༊⏬ࡢ୰࡛ືⓗ┦స⏝ࢆ⧞ࡾ㏉ࡋ࡚࠸ ࡿࡇࢆ♧၀ࡋ࡚࠾ࡾࠊᡃࠎࡀ௨๓ሗ࿌ࡋࡓࠊ⫥ ⱆ࢚ࣥࣁࣥࢧ࣮ Shh ࣉ࣮ࣟࣔࢱ࣮ࡣ᩿⥆ⓗ࡞┦ స⏝ࢆ⧞ࡾ㏉ࡍ࠸࠺ࢹ࣮ࢱࢆࡼࡃᨭᣢࡋ࡚࠸ࡓࠋ ࡉࡽࠊShh ㏆ഐࡢ Lmbr1 ࡢࣉ࣮ࣟࣔࢱ࣮ࡣࠊ Shh ẚ࡚ࡼࡾᑠࡉ࡞┦స⏝ࢻ࣓ࣥࢆᙧᡂࡋ ࡚࠾ࡾࠊShh ⩻ヂ㡿ᇦࡢ┦స⏝ࡣࢇㄆࡵ ࡽࢀ࡞ࡗࡓࠋ࣐࢘ࢫ⫥ⱆ࠾࠸࡚ࠊShh Lmbr1 ࡣ␗࡞ࡿⓎ⌧ࣃࢱ࣮ࣥࢆ࿊ࡍࡿࡓࡵࠊ୧⪅ࡢࣉࣟࣔ ࣮ࢱ࣮㡿ᇦ࠾ࡅࡿⓗ࡞┦స⏝ࢻ࣓ࣥࡢᙧ ᡂࡣࠊࡑࡢⓎ⌧ࡢ㐪࠸ࢆ⏕ࡳฟࡍࡢᚲせ࡛࠶ࡿྍ ⬟ᛶࡀ࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ChIP-seq ࡛᫂ࡽ࡞ࡗࡓ CTCF ⤖ྜࢧࢺࡀ Shh ࡢⓎ⌧ᅄ⫥ࡢ⾲⌧ᆺࡢࡼ࠺ᙳ㡪ࢆ࠼ ࡿࡢㄪࡿࡓࡵࠊCRISPR/Cas9 ⣔ࢆ⏝࠸࡚ࠊࡑ ࢀࡒࢀࣀࢵࢡ࢘ࢺࡋࡓ࣐࢘ࢫᰴࢆస〇ࡋࡓࠋ⌧ᅾࠊ ࡇࢀࡽࡢ࣐࢘ࢫࡢ⾲⌧ᆺゎᯒࡀ㐍⾜୰࡛࠶ࡿࠋ Shh ࡑࡢ㏆ഐࡢ Lmbr1 㑇ఏᏊࡣࠊ␗࡞ࡿࢧࢬ ࡢ┦స⏝ࢻ࣓ࣥࢆ᭷ࡋ࡚࠸ࡓࠋࡇࢀࡣࠊ࢚ࣥࣁ ࣥࢧ࣮㑇ఏᏊࡢ┦ᑐⓗ⨨㛵ಀ㸦ࢺ࣏ࣟࢪ࣮㸧ࡀࠊ ㏆᥋ࡍࡿ㑇ఏᏊ㛫ࡢ␗࡞ࡿⓎ⌧ࣃࢱ࣮ࣥࢆ⏕ࡳฟࡍ ྍ⬟ᛶࢆ♧၀ࡋ࡚࠸ࡿࠋ㑇ఏᏊ㛫ࡢࢺ࣏ࣟࢪ࣮ࡢ㐪 ࠸ࢆ⏕ࡳฟࡍ᪂つࢩࢫ㓄ิࢆྠᐃࡍࡿࡓࡵࠊ CRISPR/Cas9 ⣔ࢆ⏝࠸࡚ࠊShh 㑇ఏᏊᗙ㛗㊥㞳Ḟ ኻࢆ᭷ࡍࡿ」ᩘࡢ࣐࢘ࢫ⬇ࢆస〇ࡋࡓࠋࡇࢀࡽࡢ࣐ ࢘ࢫࡢⓎ⌧ゎᯒࡼࡾࠊLmbr1 ୗὶࡢ 60 Kb ࡀࠊ ᪂つࣥࢩ࣮ࣗࣞࢱ࣮ࢆྵࡴࡇࢆⓎぢࡋࡓࠋ ⌧ᅾࠊ௨ୖࡢᡂᯝࢆྵࡴㄽᩥࢆసᡂ୰࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 382 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࣐࢘ࢫ⾲⌧ᆺᐤࡍࡿࣞࣝⓎ⌧㔞ࣂࢫࡢཎᅉࢩࢫไᚚ㓄ิ ᇶ┙◊✲㸦㸿㸧 ᇛ▼ ಇᙪ࣭ሗ࣭ࢩࢫࢸ࣒◊✲ᶵᵓᅜ❧㑇ఏᏛ◊✲ᡤ ⣔⤫⏕≀◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ⏕≀✀ෆࡢ⾲⌧ᆺከᵝᛶࡢཎᅉࡢከࡃࡀࠊ㑇ఏᏊ Ⓨ⌧㔞ࢆไᚚࡍࡿࢩࢫ㓄ิከᆺ࡛࠶ࡿࡇࡀศࡗ ࡚ࡁࡓࠋࡋࡋࠊࢩࢫ㓄ิከᆺࢆయ⣔ⓗྠᐃࡍࡿ ᪉ἲࡣᮍᩚഛ࡛ࠊ⾲⌧ᆺከᵝᛶࢆ⏕ࡴࢤࣀ࣒ከᆺࡢ యീࡣᮍ▱࡛࠶ࡿࠋᮏ◊✲࡛ࡣࠊ᭱ึ࣐࢘ࢫ⾲ ⌧ᆺከᵝᛶࡢཎᅉ࡞ࡿ㑇ఏᏊ⩌ࢆᢳฟࡍࡿࡓࡵࠊ 㣖ヂ㓄ิୖࡢከᆺ(cSNP)ࢆᣦᶆࠊ࣐࢘ࢫள✀㛫 㓄 F1 ಶయࢆ⏝࠸࡚ᑐ❧㑇ఏᏊⓎ⌧㔞ẚࡢṍࡳ(ࣞ ࣝⓎ⌧㔞ࣂࢫ)ࢆィ ࡍࡿࠋḟࠊⓎ⌧㔞ࣂ ࢫ┦㛵ࡍࡿࢡ࣐ࣟࢳࣥಟ㣭ࡸᰁⰍయ࣮ࣝࣉࡢள✀ 㛫ᕪࢆᇶ㛵㐃ࡍࡿࢩࢫ㓄ิࢆ᥈⣴ࡋࠊ㔝⏕⏤᮶⣔ ⤫ࡢࣁࣉࣟࢱࣉゎᯒࠊᇵ㣴⣽⬊ࡸ࣐࢘ࢫಶయࡢ 㑇ఏᏊᑟධᐇ㦂ࡼࡾཎᅉࢩࢫከᆺࢆྠᐃࡍࡿࠋᮏ ᖺᗘࡣࠊ㌟㛗ࡸ㛗㦵㛗࡞࣐࢘ࢫள✀㛫ࡢ㢧ⴭ࡞ᙧ ែከᵝᛶࡢཎᅉ㑇ఏᏊ㸦⩌㸧ࢆ᥈⣴ࡍࡿࡓࡵࠊすḢ ⏘ Mus musculus domesticus ள✀⏤᮶ࡢ C57BL/6J ⣔⤫ࠊ᪥ᮏ⏘ Mus musculus molossinus ள✀⏤᮶ࡢ MSM/Ms ⣔⤫ࠊ࠾ࡼࡧࡇࢀࡽࡢṇ㏫㓄 F1 ಶయࢆ ⏝ࡋ࡚ࠊᡂ㛗㌾㦵ࢆᑐ㇟ࣞࣝⓎ⌧㔞ࣂࢫ ィ ࠊ࠾ࡼࡧ࣮࢜ࣉࣥࢡ࣐ࣟࢳࣥᵓ㐀ࡢ᥈⣴࣭ྠᐃ ◊✲ࢆィ⏬ࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᮏᖺᗘࡣࠊࡲࡎᡂ㛗㌾㦵ࢆᑐ㇟ḟୡ௦ࢩ࣮ࢣࣥ ࢧࢆ⏝࠸ࡓ RNA-seq ゎᯒࡢᨭࢆཷࡅࡓࠋゎᯒ⣔⤫ ࡣࠊ࣐࢘ࢫ C57BL/6J ⣔⤫ MSM/Ms ⣔⤫ࠊࡇࢀࡽ ࡢṇ㏫㓄ࡼࡾᚓࡽࢀࡓ 2 ✀㢮ࡢ F1ࠊ࠾ࡼࡧࢥࣥ ࢺ࣮ࣟࣝࡋ࡚ࡢࣛࢵࢺࡢィ 5 ✀㢮࡛࠶ࡿࠋゎᯒࡣ ࣞࣝⓎ⌧㔞ࣂࢫࡢᛶᕪࡘ࠸࡚ࡶ᳨ドࡍࡿࡓ ࡵࠊ㞤㞝୧᪉ࡘ࠸࡚⾜ࡗࡓࠋゎᯒᩘࡣ㞤㞝ྛ 6 ༉ ⏤᮶ࡢ⤌⧊ࢆࣉ࣮ࣝࡋࡓィ 10 ✀࡛࠶ࡿࠋ ࡘࡂࠊⓎ⌧㔞ࣂࢫࢆ♧ࡍ㑇ఏᏊࡢࢩࢫㄪ⠇ 㓄 ิࡢᵓ 㐀ከᆺࢆ᫂ ࡽ ࡍࡿࡓࡵ ࠊ Transposase-Accessible Chromatin (ATAC)-seq ࡼࡿ࣮࢜ࣉࣥࢡ࣐ࣟࢳࣥᵓ㐀ࡢ᥈⣴࣭ྠᐃࢆ⾜࠺ࡇ ࢆணᐃࡋ࡚࠸ࡿࡀࠊATAC-seq ࡣࡈࡃ᭱㏆ࡢᢏ⾡࡛ ࠶ࡿࡓࡵࠊࡲࡎࡣࢸࢫࢺࢧࣥࣉࣝࢆ⏝࠸࡚ᑠつᶍゎ ᯒࢆ⾜࠸ࠊᚓࡽࢀࡓ⤖ᯝࢆཧ⪃ࡋ࡚ᮏ␒ࢧࣥࣉࣝ ࡢㄪᩚἲ࡞ࢆ᳨ウࡍࡿᚲせࡀ࠶ࡿุ᩿ࡋࡓࠋࡑ ࡢࡓࡵࡢࢸࢫࢺࢧࣥࣉࣝࡣࠊᮏሗ࿌᭩సᡂⅬ(2015 ᖺ 12 ᭶)࡛ᨭ⌜㏦῭ࡳ࡛࠶ࡿࠋ⤖ᯝࡢ 2 ḟゎ ᯒࢆ⾜࠸ḟ➨ࠊᡂ㛗㌾㦵ࡽ᥇ྲྀࡋࡓࢧࣥࣉࣝࡢゎ ᯒࢆᨭ⌜౫㢗ࡍࡿࠋATAC-seq ゎᯒࡘ࠸࡚ࡶࠊ ࢧࣥࣉࣝࡣ C57BL/6J ⣔⤫ࠊMSM/Ms ⣔⤫ࠊࡇࢀࡽ ࡢṇ㏫㓄⏤᮶ࡢ 2 ✀㢮ࡢ F1ࠊ࠾ࡼࡧࣛࢵࢺࡢ 5 ✀ ⏤᮶ࡍࡿࡶࡢࢆணᐃࡋ࡚࠸ࡿࠋ ࣛࢵࢺࡢィ 5 ✀ࡢ㞤㞝ྛ 6 ༉ࡢࣉ࣮ࣝࢧࣥࣉࣝࠊィ 10 ✀㢮ࡘ࠸࡚ࠊᡂ㛗㌾㦵ࢆᑐ㇟ḟୡ௦ࢩ࣮ࢣࣥ ࢧࢆ⏝࠸ࡓ RNA-seq ࡢゎᯒ⤖ᯝ࡛࠶ࡾࠊ36pb ࢩࣥ ࢢࣝㄞࡳࠊ ᖹᆒ 3,000 ࣮ࣜࢻࡢゎᯒ⤖ᯝࢆ㡬࠸ࡓࠋ ࡇࢀࡲ࡛ࠊ࣐࢘ࢫ࠾ࡼࡧࣛࢵࢺࡢྛ⣔⤫࠾ࡼࡧ 㓄⩌ࢆ⏝࠸࡚ࠊࣞࣝⓎ⌧㔞ࣂࢫࢡ࣐ࣟࢳࣥ ಟ㣭ࡸ࣮࢜ࣉࣥࢡ࣐ࣟࢳࣥ㒊࡞ࡢ࢚ࣆࢤࣀ࣒ᵓ 㐀ከᆺࢆ㛵㐃࡙ࡅ࡚ゎᯒࡋࠊࢤࣀ࣒࣡ࢻࢩࢫ㓄 ิࢆྠᐃࡋࡓሗ࿌ࡣࡲࡔ࡞࠸ࠋ⌧ᅾࠊᨭ࠸ࡓࡔ࠸ ࡓࢹ࣮ࢱࡣ 2 ḟゎᯒ୰࡛࠶ࡿࡀࠊᮏ◊✲࡛ゎᯒࡋ࡚ ࠸ࡿࣞࣝⓎ⌧㔞ࣂࢫࠊࢩࢫ㓄ิከᆺࠊࡑࡋ࡚ ⾲⌧ᆺࡢ୕⪅ࢆయ⣔ⓗ㛵㐃࡙ࡅ࡚⾜࠺ゎᯒࡣࠊᐇ 㝿㔞ⓗᙧ㉁ᐤࡍࡿ㑇ఏᇶ┙ࡢゎ᫂ࡁࡃ㈉ ⊩ࡍࡿࡶࡢᮇᚅࡉࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᅜෆእ࡛⾜ࢃࢀࡿ࣐࢘ࢫ F1 㞧✀ࢆ⏝࠸ࡓ㑇ఏᏊ Ⓨ⌧ゎᯒࡢከࡃࡣࠊ∗⣔࣭ẕ⣔ࡢࣥࣉࣜࣥࢺຠᯝ ࢆㄝ᫂ࡍࡿࡶࡢ೫ࡗ࡚࠸ࡿࠋ୍᪉ࠊᮏ◊✲࡛ࡣࠊ C57BL/6J ⣔⤫ MSM/Ms ⣔⤫ࡢṇ㏫㓄࡛∗⣔࣭ ẕ⣔ࡢ␗࡞ࡿ 2 ✀㢮ࡢ F1 ࢆస〇ࡋࠊᪧ᮶ࡢࣥࣉࣜ ࣥࢺ㑇ఏᏊ⩌ࡳࡽࢀࡿࣞࣝⓎ⌧ࣂࢫࢆ࣏ࢪ ࢸࣈࢥࣥࢺ࣮ࣟࣝࡋ࡚ཧ⪃ࡋ࡞ࡀࡽࠊࣥࣉ ࣜࣥࢸࣥࢢ⌧㇟ࡣ↓㛵ಀ࡞᪂ࡓ࡞ள✀࠾ࡼࡧ✀ 㛫ࡢࣞࣝⓎ⌧ࣂࢫὀ┠ࡋ࡚࠸ࡿࠋࡲࡓࠊ ATAC-seq ࡼࡿ࣮࢜ࣉࣥࢡ࣐ࣟࢳࣥሗࡣࠊᡂ㛗㌾ 㦵࡛Ⓨ⌧ࡍࡿ㑇ఏᏊࡢࢩࢫㄪ⠇㓄ิࡢ⨨ሗࢆ ࠼ࡿࡶࡢ࡛࠶ࡿࠋᚋࠊࣞࣝⓎ⌧ࣂࢫ࣮࢜ ࣉࣥࢡ࣐ࣟࢳࣥࡢ⛬ᗘࡢࣞࣝ㛫ᕪ㐪ࢆᑐᛂ࡙ࡅࡿ ࡇࡀ࡛ࡁࢀࡤࠊ᪂つ࡞ࢩࢫ㓄ิྠᐃἲࡘ࡞ࡆ ࡿࡇࡀྍ⬟࡞ࡿࠋ ࡉࡽࠊ࣐࢘ࢫள✀㛫࠾ࡅࡿࣞࣝⓎ⌧㔞ࣂ ࢫࡣࠊ㦵ࢧࢬ࡞㔞ⓗᙧ㉁ࡢள✀㛫㸦✀㛫㸧ከ ᵝᛶࢆㄝ᫂ࡍࡿせ࡞㑇ఏᇶ┙࡞ࡾ࠺ࡿࠋࡲࡓࠊ ᮏࢤࣀ࣒ゎᯒᨭࡼࡗ࡚⏕⏘ࡉࢀࡿࢹ࣮ࢱࡣࠊᙧ ែከᵝᛶࡢཎᅉ࡞ࡿࢩࢫไᚚ㓄ิࡢᇶ♏࡞ࡿࡶ ࡢ࡛࠶ࡾࠊᙧែ㐍ࡢ㑇ఏᇶ┙ࡢ᰿ᖿࡢ⌮ゎࡘ࡞ ࡀࡿࡶࡢ࡛࠶ࡿࠋࡇࢀࡽࡢሗࡇࢀࡲ࡛ᡃࠎࡀ ✚ࡋ࡚ࡁࡓ⾲⌧ᆺሗࢆ㛵㐃࡙ࡅࡿࡇ࡛ࠊࢩ ࢫไᚚ㓄ิከᆺࡀ㔞ⓗᙧ㉁ࡢள✀㛫㸦✀㛫㸧ከᵝᛶ ࢆ⏕ࡳฟࡍ࣓࢝ࢽࢬ࣒ࡢయീࢆ᫂ࡽࡍࡿࡇ ࡀྍ⬟࡞ࡿ⪃࠼࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 㑇ఏᏊⓎ⌧ゎᯒࡢࢹ࣮ࢱࡣࠊDDBJ Ⓩ㘓‽ഛ୰ ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏሗ࿌᭩సᡂⅬ࡛ࠕࢤࣀ࣒ᨭࠖࡼࡾཷࡅࡓ ⤖ᯝࡣࠊ࣐࢘ࢫ C57BL/6J ⣔⤫ MSM/Ms ⣔⤫ࠊࡇ ࢀࡽࡢṇ㏫㓄ࡼࡾᚓࡽࢀࡓ 2 ✀㢮ࡢ F1ࠊ࠾ࡼࡧ - 383 - බເᨭㄢ㢟㸸2015 ᖺᗘ ⭠⟶ ,J$ ᢠయࡼࡿ⭠ෆ⣽⳦㑅ูᶵᵓࡢゎ᫂ ᇶ┙◊✲㸦㹀㸧 ᪂ⶶ ♩Ꮚ࣭㛗ࣂ࢜Ꮫࣂ࢜ࢧ࢚ࣥࢫᏛ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ⭠⟶ෆࡣከ✀ከᵝ࡞⣽⳦ࡀᖖ⏕ᜥࡋࠊᐟ ᖹⓗ࡞ඹ⏕㛵ಀࢆ⠏࠸࡚࠸ࡿࠋࡇࡢඹ⏕㛵ಀࡀᔂ ࢀࡿࠊ⅖ᛶ⭠ᝈࡸ⫧‶ࠊ⢾ᒀࢆࡣࡌࡵࡍ ࡿ⏕ά⩦័ࠊ⭠࢞ࣥ࡞ྛ✀ᝈࡢⓎ⧅ࡀ ࡿࡓࡵࠊ⭠ෆ⎔ቃࢆᜏᖖⓗ⥔ᣢࡍࡿࡇࡣᗣ⥔ ᣢ㔜せ࡛࠶ࡿࠋ⭠ࡣᐟ⭠ෆ⣽⳦ࡼࡿඹ⏕ࡢ 㔜せ࡞ሙ࡛࠶ࡾࠊᜏᖖᛶ⥔ᣢࡢࡓࡵከࡃࡢච⣽ ⬊ࡸศᏊࡀാ࠸࡚࠸ࡿࡀࠊIgA ࡣ⢓⭷㠃ࡢཎ⳦㜵 ᚚࡔࡅ࡛࡞ࡃ⭠ෆᖖᅾ⣽⳦ࡢไᚚࡶ㔜せ࡛࠶ࡾࠊ ࡇࡢࡼ࠺࡞ඹ⏕㛵ಀࡢ⥔ᣢࡶᴟࡵ࡚㔜せ࡛࠶ࡿ ⪃࠼ࡽࢀ࡚࠸ࡿࠋ⭠⟶ศἪࡉࢀࡿ IgA ࡣ⭠ෆ⣽⳦ ᑐࡋ࡚ poly-reactive ᛂࡋ⢓⭷㠃ࡢ➨୍⥺㜵ᚚ 㔜せ⪃࠼ࡽࢀ࡚ࡁࡓࡀࠊලయⓗ⭠ෆ⣽⳦ࢆ ࡢࡼ࠺ㄆ㆑ࡋไᚚࡍࡿࡣ᫂ࡽ࡛ࡣ࡞࠸ࠋ᭱㏆ ࡢ◊✲࡛ࡣ⭠⟶⭍ศἪࡉࢀࡿ IgA ᢠయࡀ⭠ෆ⣽⳦ ࢆ㆑ูࡋ࡚ไᚚࡍࡿࡇࡀ♧၀ࡉࢀ࡚࠸ࡿࡀලయⓗ ࡞ࢱ࣮ࢤࢵࢺࡣ᫂ࡽࡉࢀ࡚࠸࡞࠸ࠋ ⚾ࡓࡕࡣ⭠⟶ࡢᜏᖖᛶࢆ⥔ᣢࡍࡿࡓࡵࡢࣔࣀࢡࣟ ࣮ࢼࣝ IgA ᢠయ㸦W27㸧ࢆ࣐࢘ࢫࡽ༢㞳ࡋࠊࡑࡢ ࢱ࣮ࢤࢵࢺศᏊ࢚ࣆࢺ࣮ࣉࡶྠᐃࡋࡓࠋW27 ᢠయ ࡀㄆ㆑ࡍࡿศᏊࡣࡰࡍ࡚ࡢ⣽⳦ࡼࡗ࡚Ⓨ⌧ࡉ ࢀ࡚࠸ࡿࡀࠊ࢚ࣆࢺ࣮ࣉ㒊ศࡢ࣑ࣀ㓟㓄ิࡣ⣽⳦ ࡈᅛ᭷࡛࠶ࡾࠊW27 ᢠయࡀ࢚ࣆࢺ࣮ࣉ㒊ศࡢ ࣑ࣀ㓟㓄ิࡢᕪ␗ࢆ༊ูࡋ࡚ㄆ㆑ࡍࡿࡇࡶ᫂ࡽ ࡋࡓࠋࡇࡢᢠయࢆ࣐࢘ࢫ⤒ཱྀᢞࡍࡿࡇ࡛ࠊ ⭠ෆ⣽⳦ྀࡢࣂࣛࣥࢫࡀኚࡋࠊ࣐࢘ࢫ⭠⅖ࣔࢹࣝ ࡛ࡣ W27 ᢠయ⤒ཱྀᢞࡼࡿ⭠⅖ᢚไຠᯝࡶ☜ㄆ ࡋࡓࠋ㸦ㄽᩥᢞ✏‽ഛ୰㸧 ᮏ◊✲ᥦࡢ┠ⓗࡣࠊᐟࡢ IgA ࡀࡑࡢᢠཎ≉␗ ᛶࢆࡗ࡚ከ✀ከᵝ࡞⭠ෆ⣽⳦ࡢඹ㏻ㄆ㆑ศᏊࡢ࢚ ࣆࢺ࣮ࣉࡢᕪ␗ࢆ㏻ࡋ࡚ྲྀᤞ㑅ᢥࢆࡍࡿࡇ࡛ඹ⏕ 㛵ಀࢆಖᣢࡍࡿࠊ࠸࠺ᡃࠎࡢ௬ㄝࢆศᏊ࡛ࣞ࣋ࣝ ド᫂ࡍࡿࡇ࡛࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ⚾ࡓࡕࡀ࣐࢘ࢫ⭠⟶ࡽྲྀᚓࡋࡓ W27 IgA ᢠయࡣࠊ Lactobacillus casei ࡸ Bifidobacterium bifidum ࡢ ࡼ࠺࡞࠸ࢃࡺࡿၿ⋢⳦ࡣ⤖ྜࡏࡎཎ⳦ࢆྵࡴ ࡢከࡃࡢ⣽⳦ࡣ⤖ྜࡋ࡚⭠ෆ⣽⳦ྀࢆⰋ࠸≧ែ ኚࡉࡏࡿணࡉࢀࡿᢠཎ≉␗ᛶࢆᣢࡗࡓᢠయ࡛ ࠶ࡿࠋW27 ᢠయࡀ⤖ྜࡍࡿࠊ࠶ࡿ࠸ࡣ⤖ྜࡋ࡞࠸⣽ ⳦⩌ࡑࢀࡽࡢ⣽⳦ࡢ௦ㅰᶵ⬟ࢆ᫂ࡽࡍࡿࡇ ࡀ࡛ࡁࢀࡤࠊᐟࡀ IgA ᢠయࢆ㥑ࡋ࡚┠ᣦࡋ࡚࠸ ࡿⰋ࠸⭠ෆ⎔ቃࡀ࠸࡞ࡿ≧ែࠊࢆᐃ⩏ࡍࡿࡇ ࡀ࡛ࡁࡿᛮ࠺ࠋᮏᨭ࡛ࡣ௨ୗࡢࡘࡢᐇ㦂ࢧࣥ ࣉࣝࡢ࣓ࢱࢤࣀ࣒ゎᯒ࣭࣓ࢱ 16S rRNA ゎᯒࢆ⾜ࡗ ࡓࠋ 㸯ࠊW27 ᢠయᢞᐇ㦂๓ᚋࡢ⣅౽⏤᮶ࡢ DNA ࢆ ࡗ࡚ 16S rRNA ゎᯒࢆ⾜࠸ࠊ⭠ෆ⣽⳦ྀࡀ W27 ᢞ ࡼࡾ᭷ពኚࡍࡿࢹ࣮ࢱࢆࡍ࡛ᚓ࡚࠸ࡿࠋᮏ ᨭ࡛ࡣྠࡌࢧࣥࣉࣝࢆ⏝࠸࡚ࠊ⭠ෆ⣽⳦ྀࡢ࣓ࢱ ࢤࣀ࣒ゎᯒࢆ⾜࠸ࠊᢠయࡢ⤒ཱྀᢞ๓ᚋࡢ⭠ෆ⎔ቃ ࡢ㐪࠸ࢆ⣽⳦ࡢ✀㢮ࡔࡅ࡛ࡣ࡞ࡃᚤ⏕≀ࡢᶵ⬟࠸ ࠺ほⅬࡽヲ⣽᳨ウࡍࡿࠋW27 ᢞ࣐࢘ࢫ 㸲༉ ࡢᢠయᢞ๓ᚋࠊィ 8 ࢧࣥࣉࣝࡢ࣓ࢱࢤࣀ࣒ゎᯒࢆ ᨭ࠸ࡓࡔ࠸ࡓࠋ 㸰ࠊ⭠⟶ෆᐜ≀ࢆ IgA ᢠయᑐࡍࡿᢠయ࡛ᰁⰍࡋ࡚ IgA ⤖ྜ⣽⳦⩌ IgA 㠀⤖ྜ⣽⳦⩌ࢭࣝࢯ࣮ࢱ࣮ ࡛ศ㞳ࡍࡿࡀྍ⬟࡛࠶ࡿࠋࡇࡢ᪉ἲࢆᛂ⏝ࡋ࡚ࠊ W27 IgA ᢠయࡼࡿ⭠ෆ⣽⳦ྀไᚚࡢ㑅ᢥᇶ‽ࢆ᫂ ࡽࡍࡿࡇࡀ࡛ࡁࡿ⪃࠼ࡿࠋලయⓗࡣࠊIgA ࡀࡃ⭠⟶ෆᏑᅾࡋ࡞࠸ AID ࣀࢵࢡ࢘ࢺ࣐࢘ࢫ ࡢ┣⭠ෆᐜ≀ࢆ W27 ᢠయ࡛ᰁⰍࡍࡿࠋࢭࣝࢯ࣮ࢱ࣮ ࡛ W27 ᢠయ⤖ྜ⣽⳦ W27 ᢠయ㠀⤖ྜ⣽⳦ศ㞳 ࡋࠊDNA ࢆᢳฟᚋ 16S rRNA ゎᯒ࣓ࢱࢤࣀ࣒ゎ ᯒࢆ⾜࠺ࠋAID ࣀࢵࢡ࢘ࢺ࣐࢘ࢫ㸲༉ࡽࢧࣥࣉ ࣝࢆᢳฟࡋࠊィ㸶ࢧࣥࣉࣝࡑࢀࡒࢀࡢ࣓ࢱࢤࣀ࣒ゎ ᯒ 16S rRNA ゎᯒࡢᨭࢆ࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣓ࢱࢤࣀ࣒ゎᯒࠊ࠾ࡼࡧ࣓ࢱ 16S rRNA ゎᯒࡣ㐍⾜ ୰࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ゎᯒ⤖ᯝࡀᮍࡔ࡞࠸ࡓࡵࠊヱᙜ࡞ࡋࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ゎᯒ⤖ᯝࡀᮍࡔ࡞࠸ࡓࡵࠊヱᙜ࡞ࡋࠋ - 384 - බເᨭㄢ㢟㸸2010㹼2011 ᖺᗘ ࢮ ࣈࣛࣇࢵࢩࣗ㏆⣔ࡢᶞ❧㸭ࢮࣈࣛࣇࢵࢩࣗึᮇ⢭ཎ⣽⬊␗ᖖኚ␗యࡽࡢ⢭ཎᖿ ⣽⬊ቑṪ࣭ศᅉᏊࡢྠᐃ ⱝᡭ◊✲㸦㹀㸧㸭᪂Ꮫ⾡㡿ᇦ◊✲ ᪂ᒇ ࡳࡢࡾ࣭ᅜ❧㑇ఏᏛ◊✲ᡤ⣔⤫⏕≀◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ᑠᆺ㨶㢮࡛࠶ࡿࢮࣈࣛࣇࢵࢩࣗࡣ㢖⦾⏝ࡉࢀ ࡚࠸ࡿࣔࢹࣝື≀ࡢ୍ࡘ࡛࠶ࡿࡀࠊ㛗ᖺࠊ㑇ఏⓗ ᆒ୍࡞㏆⣔ࡀᶞ❧ࡉࢀ࡚ࡇ࡞ࡗࡓࠋࡑࡇ࡛ᨭ ౫㢗⪅ࡣࠊ2005 ᖺࡼࡾࢮࣈࣛࣇࢵࢩࣗࡢ㔝⏕ᆺ⣔ ⤫࡛࠶ࡿ India ࢆ⏝࠸ࠊ࣓ࢲ࢝ࡸ࣐࢘ࢫ࡛ࡢ㏆⣔ ࡢᐃ⩏࡛࠶ࡿ 20 ୡ௦௨ୖࡢጒ㓄ࢆヨࡳࡓࠋ㏵୰ࠊ ⥅௦ࡀ㞴ࡋ࠸ᮇࡀ࠶ࡗࡓࡶࡢࡢࠊ2010 ᖺ㸦ᨭ౫ 㢗㸧ࡣ 16 ୡ௦┠ࡢಶయࢆᚓࡿࡇࡀ࡛ࡁࠊ2011 ᖺࡣ 20 ୡ௦┠฿㐩ࡋࡓࠋIM㸦India-Mishima㸧 ⣔⤫ྡࡅࡓࡇࡢ⣔⤫ࡣࠊୡ⏺࡛ึࡵ࡚ࡢࢮࣈࣛ ࣇࢵࢩࣗ㏆⣔࡛࠶ࡗࡓࠋ ⥅௦㏵୰ࡢࢥࣥࢱ࣑ࢿ࣮ࢩࣙࣥࡣ༑ศὀពࢆᡶ ࡗ࡚࠸ࡓࡀࠊࡑࡢྍ⬟ᛶࢆ᫂☜㝖ࡍࡿᚲせࡀ࠶ ࡿࠋࡲࡓࠊIM ⣔⤫ࡢ⏝ࢆ⪃࠼ࡓࡁࠊࡇࡢ⣔⤫ෆ ࡢ⛬ᗘࡢከᆺࡀṧࡗ࡚࠸ࡿࡢࠊࡑࡋ࡚ࡑࡢከ ᆺࡣࢤࣀ࣒ୖࡢࡇ࠶ࡿࡢࢆᢕᥱࡋ࡚࠾ࡃࡇ ࡣ㔜せ࡛࠶ࡿࠋࡉࡽࠊᶞ❧ᡂຌࡋࡓ㏆⣔ࢆ⮬ ศ⮬㌟ࡔࡅ࡛ࡣ࡞ࡃࠊୡ⏺୰ࡢ◊✲⪅ᗈࡃ⏝ࡋ ࡚ࡶࡽ࠺ࡓࡵࡣࠊ⏝ࡋࡸࡍ࠸ᙧ࡛ሗࢆබ㛤ࡍ ࡿࡇࡀษࡔ⪃࠼ࡓࠋࡑࡇ࡛ࠊ௨ୗ㏙ࡿ୍ 㐃ࡢᨭࢆ౫㢗ࡋࡓࠋ ⣔⤫ࢆᇶసᡂࡉࢀࡓࢮࣈࣛࣇࢵࢩࣗබ㛤ࢤࣀ ࣒㓄ิࡢ㛫ࡢ⛬ᗘࡢከᆺࡀᏑᅾࡍࡿࡢࢆ▱ ࡿࡇࡀ࡛ࡁࡿࠋ 14 ୡ௦┠ࡢ 2 ಶయ㛫ࡢ୍ሷᇶከᆺࡣ 0.14㸣⛬ᗘ࡛ ࠶ࡗࡓࡇࡽࠊࢥࣥࢱ࣑ࢿ࣮ࢩࣙࣥ➼࡞ࡃ㏆⣔ ࡀ㐍⾜࡛ࡁ࡚࠾ࡾࠊ⌧Ꮡࡢ IM ⣔⤫ෆࡣ 0.14㸣 ௨ୗࡢከᆺࡋᏑᅾࡋ࡞࠸ࡇࡀ᥎ᐃ࡛ࡁࡓࠋ࣐࢘ ࢫ➼ࡢࢹ࣮ࢱ↷ྜࡋ࡚ࡶࠊ㏆⣔ࡋ࡚༑ศᆒ ୍࡛࠶ࡿゝ࠼ࡿࠋࡲࡓࠊබ㛤㓄ิࡢ㛫ࡣ 1㸣⛬ ᗘࡢ୍ሷᇶ⨨ࡀᏑᅾࡋ࡚࠸ࡓࠋᚑࡗ࡚ࠊࢮࣈࣛࣇ ࢵࢩࣗࡣ⣔⤫㛫ከᆺᐩࢇ࡛࠾ࡾࠊ㑇ఏᏛⓗ࡞ゎ ᯒࢆ⾜࠺Ⰻ࠸ࢶ࣮࡛ࣝ࠶ࡿࡇࡀᨵࡵ࡚☜ㄆ࡛ࡁࡓࠋ ᮏᨭࡼࡾసᡂࡉࢀࡓࢹ࣮ࢱ࣮࣋ࢫࡣࠊIM ⣔⤫ ࢆ⏝࠸ࡓᵝࠎ࡞㡰㑇ఏᏛⓗ࠾ࡼࡧ㏫㑇ఏᏛⓗゎᯒࢆ 㐍ࡵࡿࡇࢆྍ⬟ࡍࡿࡶࡢ࡛࠶ࡿࠋ㏆ᖺࠊ ࢮࣈࣛࣇࢵࢩࣗ࠾ࡅࡿ㏫㑇ఏᏛⓗᡭἲࡋ࡚ TALEN 㸦 Transcription Activator-Like Effector Nucleases 㸧 ࡸ CRISPR 㸦 Clustered Regularly Interspaced Short Palindromic Repeats㸧ࢆ⏝࠸ࡓ ࢤࣀ࣒⦅㞟ᢏ⾡ࡀᛴ㏿ᗈࡲࡾࡘࡘ࠶ࡿࠋࡇࢀࡽࡢ ゎᯒ࠾࠸࡚ᙜヱࢹ࣮ࢱ࣮࣋ࢫࡣά⏝ࡍࡿࡇࡼ ࡾࠊゎᯒࡢຠ⋡ࢆᅗࡿࡇࡀ࡛ࡁࡿࠋ 㸺ᨭࡢෆᐜ㸼 ḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮ࡼࡿࢤࣀ࣒㓄ิỴᐃ㸦2010 ᖺᗘ㸧ࠊ࠾ࡼࡧࡑࡢࢹ࣮ࢱࢆ㑇ఏᏛⓗゎᯒ⏝ࡍࡿ ࡓࡵࡢࢹ࣮ࢱ࣮࣋ࢫᵓ⠏㸦2011 ᖺᗘ㸧ࡘ࠸࡚ᨭ ࢆཷࡅࡓࠋ ࢤࣀ࣒㓄ิỴᐃ⏝࠸ࡓࢧࣥࣉࣝࡣࠊጒ㓄 14 ୡ௦┠ࡢ IM ⣔⤫ 2 ಶయ㸦⥅௦࣌㸧࡛࠶ࡗࡓࠋᚓ ࡽࢀࡓ㓄ิࢆ᪤බ㛤ࡉࢀ࡚࠸ࡿࢮࣈࣛࣇࢵࢩࣗ ࢤࣀ࣒㓄ิᑐࡋ࡚࣐ࢵࣆࣥࢢࡋࠊබ㛤㓄ิࡢ㛫 ࡢከᆺࠊ࠾ࡼࡧ㓄ิỴᐃࡉࢀࡓ㏆⣔ 2 ಶయ㛫ࡢከ ᆺࡶ᳨ฟ࣭㞟ィࡋ࡚㡬࠸ࡓࠋ᭦ࠊᚋᵝࠎ࡞㑇ఏ Ꮫⓗゎᯒ࡚ IM ⣔⤫ࢆ᭷ຠά⏝ࡍࡿࡓࡵࠊ᪤▱ 㓄ิ࠾ࡼࡧከᆺሗࠊIM ⣔⤫ࡢ㓄ิሗࠊIM ⣔⤫ ᪤▱㓄ิ㛫ࡢከᆺሗ࡞ࢆྵࡵࡓࢤࣀ࣒㛵㐃 ሗࢆ㞟⣙ࡋࡓࢹ࣮ࢱ࣮࣋ࢫࠊ࠾ࡼࡧࡑࡢሗࢆ⏝ ࡋࡸࡍࡃࡍࡿࡓࡵࡢࣅ࣮࣮ࣗ࣡ࡢసᡂᨭࢆ⾜ࡗ࡚ 㡬࠸ࡓࠋᑦࠊࡇࡢࢹ࣮ࢱ࣮࣋ࢫᵓ⠏ᨭࡘ࠸࡚ࡣࠊ 㜰Ꮫࡢ୰㇂᫂ᘯ≉௵ᩍᤵࡢඹྠ◊✲ࡋ࡚㐍 ࡵࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 IM ⣔⤫ࡣศࡶ㛤ጞࡋ࡚࠾ࡾࠊ᪤ᅜෆእ 10 ᩘ ௳⛬ᗘ㏦ࡋ࡚࠸ࡿࠋᨭࡼࡾᚓࡽࢀࡓࢤࣀ࣒㓄 ิࡸከᆺሗࡣࠊIM ⣔⤫ຍ౯್ࢆ࠼ࡿ㔜せ࡞ ᇶ┙ሗ࡛࠶ࡿࠋ࢞ࣥ◊✲ࡸ࣓ࢳࣝ㛵ࡍࡿ◊✲ ࡞ࠊᖜࡢᗈ࠸◊✲ศ㔝ᦠࢃࡿ◊✲⪅ࡽࡢศ ౫㢗ࢆཷࡅ࡚࠾ࡾࠊᡃࠎࡀᙜึ⪃࠼࡚࠸ࡓ௨ୖከ ᵝ࡞ศ㔝ࡢⓎᒎ㈉⊩ࡍࡿࡇࡀᮇᚅ࡛ࡁࡿࠋᮏᨭ ࡼࡾᚓࡽࢀࡓࢹ࣮ࢱ࠾ࡼࡧࡑࡢࢹ࣮ࢱ࣮࣋ࢫࢆ ୍⯡බ㛤ࡋࡓࡇࡼࡾࠊIM ⣔⤫ࡢศඛ࠾࠸ ࡚ࡶά⏝࡛ࡁࠊຠ⋡ⓗ◊✲ࢆ㐍ࡵࡽࢀࡿ⪃࠼࡚ ࠸ࡿࠋ IM ⣔⤫ 2 ಶయࡑࢀࡒࢀࢮࣈࣛࣇࢵࢩࣗබ㛤ࢤ ࣀ࣒㓄ิࡢẚ㍑ࠊ࠾ࡼࡧ IM ⣔⤫ෆࡢከᆺㄪᰝ ࡼࡾࠊጒ㓄ࡼࡗ࡚ከᆺࡀኻࢃࢀࡘࡘ࠶ࡿ୰࡛ ࡶࠊ≉ච➼ࡢ⏕య㜵ᚚ㛵㐃ࡢ㑇ఏᏊ࠾࠸࡚ࡣ ከᆺࡀ⥔ᣢࡉࢀࡿࡇࡀ᫂ࡽ࡞ࡾࡘࡘ࠶ࡿࠋࡇ ࡢࡼ࠺࡞ゎᯒࢆࢤࣀ࣒య᭦ᣑࡉࡏࡿࡇ ࡼࡾࠊࢮࣈࣛࣇࢵࢩࣗࡀ✀ࡋ࡚⥔ᣢࡉࢀࡿࡓࡵ ᚲせ࡞ࢤࣀ࣒ࡢከᵝᛶࡀ࠺࠸ࡗࡓࡶࡢࢆ᥈ࢀ ࡿྍ⬟ᛶࡀฟ࡚ࡁࡓ⪃࠼࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⌧ᅾ IM ⣔⤫ࡣࠊ㏆⣔ࡢᐃ⩏ࢆ‶ࡓࡍࡔࡅࡢ⥅௦ ࢆ⾜࠸ࠊ㏆⣔ࡋ࡚ጒ㓄ࡼࡿ⥔ᣢࢆ⥆ࡅ࡚ ࠸ࡿࠋ⌧Ⅼ࡛㣫⫱ࡋ࡚࠸ࡿ࡚ࡢಶయࡀࠊ㓄ิỴ ᐃࡉࢀࡓ 2 ಶయ⏤᮶ࡍࡿࠋᚑࡗ࡚ࠊᨭࡼࡗ࡚ ᚓࡽࢀࡓከᆺሗࡼࡾࠊ㏆⣔ IM ⣔⤫ࡢ㑇ఏⓗ⫼ ᬒࡀࡢ⛬ᗘ௨ୖᆒ୍࡛࠶ࡿࢆ᥎ᐃ࡛ࡁࡿࠋࡲࡓࠊ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ヱᙜ࡞ࡋ - 385 - බເᨭㄢ㢟㸸2012 ᖺᗘ ᯤⲡ⳦⣽⬊ෆࡢ⣽⬊࿘ᮇᅇ㊰ࡢேᕤྜᡂ ᪂Ꮫ⾡㡿ᇦ◊✲ ᮎḟ ṇᖾ࣭❧ᩍᏛ⌮Ꮫ㒊⏕⌮Ꮫ⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 +LV ࠾ࡼࡧ 6WUHS ࢱࢢ⼥ྜ 'QD$ ࡢࢡ࣐ࣟࢳࣥࢱࣥ 'QD$ ࡢ &K,36HT ゎᯒ㸸 ࢹ࣒ࣇࢽࢸ࣮⢭〇&K$3ࢆ⾜ࡗࡓࠋ ⭠⳦ࢤࣀ࣒」〇㛤ጞ⺮ⓑ㉁ 'QD$ ࡣࠊ'QD$ER[ ࡤࢀࡿ ES ࡢ '1$ 㓄ิ⤖ྜࡍࡿࠋ'QD$ER[ ࡣ」〇 㛤ጞⅬ RUL& ୖࢡࣛࢫࢱ࣮ࢆᡂࡋ࡚Ꮡᅾࡍࡿࡔࡅ࡛ ࡞ࡃࠊࢤࣀ࣒ୖ⣙ ࢝ᡤᩓᅾࡋࠊ㑇ఏᏊⓎ⌧ไ ᚚࡸ」〇㛤ጞไᚚᶵ⬟ࡋ࡚࠸ࡿࠋ⛉◊㈝◊✲ㄢ㢟 ࡣ 'QD$ άᛶࡢ⣽⬊࿘ᮇ࢜ࢩ࣮ࣞࢩࣙࣥ╔┠ࡋࡓࡶ 'QD$ ࡢ $73$'3 ᆺࣞ࣋ࣝࡢ᧯సࡣࠊࢤࣀ࣒」〇㛤 ጞࡢ␗ᖖࢆࡶࡓࡽࡋࠊ⣽⬊ቑṪࢆ㜼ᐖࡍࡿࠋࡇࡢࡓ ࡵࠊGQD$RUL& Ḟኻᰴࢆ⏝ࡋࠊḟⓗ࡞ᙳ㡪ࢆ 㝖ࡍࡿᕤኵࢆࡋࡓࠋ 'QD$$73 ຍỈศゎᅉᏊ㸦άᛶᅉᏊ㸧ࡢḞᦆ ᰴ࠾ࡼࡧ㐣Ⓨ⌧ᰴ ࡢ࡛࠶ࡿࠋ'QD$ ࡢάᛶᆺ㸦$73 ⤖ྜᆺ㸧άᛶᆺ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㸦$'3 ⤖ྜᆺ㸧࡛ࠊ'QD$ER['1$ ᩿∦⮬㌟ᑐࡍ ⭠⳦」〇㛤ጞ 'QD$ ࡢࢤࣀ࣒࣡ࢻ࡞⤖ྜࣃࢱ࣮ࣥ ࡿ⤖ྜ⬟ࡣࡁࡃኚࢃࡽ࡞࠸ࠋ୍᪉࡛㏆ᖺࠊ⣽⬊࿘ ࢆᚓࡿࡀ࡛ࡁࡓࠋᚓࡽࢀࡓ⤖ྜ㡿ᇦࡣࠊࡇࢀࡲ ᮇไᚚࡉࢀࡿ࠸ࡃࡘࡢࣉ࣮ࣟࣔࢱ࣮ࡸ RUL&'1$ ࡛ಶࠎࡑࡢ⤖ྜࡀ♧ࡉࢀ࡚ࡁࡓ」〇㛤ጞ㡿ᇦࠊ ᑐࡍࡿࣇࢵࢺࣉࣜࣥࢺゎᯒࡽࠊ'QD$ ࡢ $73 ⤖ྜᆺ 'QD$ άᛶ㡿ᇦࠊ㌿ࣉ࣮ࣟࣔࢱ࣮㡿ᇦࡶྵࡲࢀ ≉␗ⓗ⤖ྜࡍࡿࡼ࠺࡞ '1$ 㒊ࡀ࠶ࡿࡀศࡗ ࡚࠾ࡾࠊࡑࡢ┦ᑐⓗ࡞⤖ྜ㢖ᗘࡢ㐪࠸ࡶ▱ࡿࡀ࡛ ࡚ࡁࡓࠋࡋࡋ࡞ࡀࡽࠊࡑࡢࢥࣥࢭࣥࢧࢫ㓄ิࡣ ࡁࡓࠋࢿ࢞ࢸࣈࢥࣥࢺ࣮࡛ࣟࣝࡶࡳࡽࢀࡿ㠀≉␗ ᫂░࡛࠶ࡾࠊࢤࣀ࣒ࢫࢣ࣮࡛ࣝࡢ⤖ྜ㡿ᇦྠᐃࡶ࡞ ⓗ࡞⤖ྜࡀከࡃ᳨ฟࡉࢀ࡚࠸ࡿ࠸࠺ၥ㢟ࡀ᭷ࡿࡶ ࡉࢀ࡚࠸࡞࠸ࠋࡑࡇ࡛ࠊ ࠕࢤࣀ࣒ᨭࠖάືࡼࡿᨭ ࡢࡢࠊᚋࠊ'QD$ ࡢ⣽⬊࿘ᮇไᚚᅉᏊࡋ࡚ࡢᙺ ࢆཷࡅࠊάᛶᆺ 'QD$㸦$73 ⤖ྜᆺ㸧άᛶᆺ 'QD$ ࢆ◊✲ࡋ࡚࠸ࡃୖ࡛ࠊ㈗㔜࡞ሗ࡞ࡿࡶࡢ࡛࠶ࡿࠋ 㸦$'3 ⤖ྜᆺ㸧࠾ࡅࡿ⭠⳦ࢤࣀ࣒㸦0E㸧ୖ ࡉࡽࠊ'QD$ άᛶࣞ࣋ࣝᛂࡌࡓ⤖ྜࣃࢱ࣮ࣥࡢኚ ࡢ⤖ྜ㒊ࢆ &K,3VHT ࡼࡾྠᐃࡋࠊࡑࡢᕪ␗ࢆ᫂ ࢆ▱ࡿ ࡓࡵ ࠊ'QD$ άᛶ ไᚚࡢ ኚ␗ ᰴ㸦ୖ グࠊ ࡽࡍࡿࡇࢆヨࡳࡓࠋ 6WUDLQࠥ6WUDLQ㸧ࢆ⏝࠸ࡓྠᵝࡢゎᯒࢆ⾜ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࡋࡋ࡞ࡀࡽࠊࡇࢀࡽࡢኚ␗ᰴ࡛ࡣ༑ศ≉␗ⓗ࡞ ௨ୗ♧ࡍኚ␗ᰴࢆ⏝ࡍࡿࡇ࡛⣽⬊ෆ⣙ ศᏊ࠶ࡿ 'QD$ ࡢ $73$'3 ⤖ྜᆺࣞ࣋ࣝࢆ᧯సࡍࡿ ࡀྍ⬟࡛࠶ࡿࠋࡇࢀࡽࡢᰴࡽ 'QD$ ࢆᅇࡋࠊ⤖ྜ ⤖ྜࣃࢱ࣮ࣥࢆᚓࡿࡀ࡛ࡁ࡞ࡗࡓࠋࡇࡕࡽࡘ ࠸࡚ࡣᚋࠊኚ␗ᰴࡢタィࡽぢ┤ࡋࠊ᳨ウࡍࡿ ᚲせࡀ࠶ࡿࠋ ࡋ࡚࠸ࡿ '1$ ࢆᅇࡍࡿࠋᅇࡉࢀࡓ '1$ ࡢ㓄ิゎ ㄞࡘ࠸࡚ࠊḟୡ௦ᆺ '1$ ࢩ࣮ࢣࣥࢧࢆ⏝࠸ࡓᨭ ⭠⳦ 'QD$ ⺮ⓑ㉁ࡣࠊᰁⰍయ」〇㛤ጞ㡿ᇦ࣓ࣥ ࢆ࠾㢪࠸ࡋࡓࠋ ⤖ྜࡍࡿࡶࡢࡢࠊࡑࢀ௨እࡢࢤࣀ࣒ୖࡢከᵝ࡞㡿 㸦ゎᯒ⏝࠸ࡓ⭠⳦ᰴࡘ࠸࡚㸧 6WUDLQ 0* 㔝⏕ᰴ㸦ࢿ࢞ࢸࣈࢥࣥࢺ࣮ࣟࣝ㸧 ᇦࡶ⤖ྜࡋ࡚࠸ࡿࡇࢆ☜ㄆࡍࡿࡀ࡛ࡁࡓࠋ 'QD$ ࡢ⣽⬊࿘ᮇไᚚᅉᏊࡋ࡚ࡢᶵ⬟ゎᯒࡣࠊ⌧ᅾࠊ 6WUDLQ6WUDLQKLVVWUHSGQD$'GQD$ ᮏᨭࡼࡗ᳨࡚ฟࡉࢀࡓࢤࣀ࣒㡿ᇦࡢ㸯ࡘࢆࢱ࣮ 6WUDLQ6WUDLQ'RUL&'UQK$ 6WUDLQ6WUDLQ'KGD >$73'QD$ ✚ኚ␗@ 6WUDLQ6WUDLQS7DTKGD 㸲 >$73'QD$ ῶᑡኚ ␗@ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢤࢵࢺࡋ࡚㐍ࡵ࡚࠸ࡿࠋᚋࠊࡇࡢ㡿ᇦࡢゎᯒࡀ 㐍ࢇࡔᚋࠊ'QD$ άᛶࣞ࣋ࣝᛂࡌࡓ⤖ྜࣃࢱ࣮ࣥ ኚືࡢࢤࣀ࣒ࢫࢣ࣮࡛ࣝࡢゎᯒᗘᣮᡓࡍࡿணᐃ ࡛࠶ࡿࠋ - 386 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࢺࣆ࣮ᛶ⓶⅖ឤᰁ㯤Ⰽࣈࢻ࢘⌫⳦ࡢཎᛶᇶ┙ ᇶ┙◊✲㸦㹁㸧 Ⳣ ᇶ⾜࣭ᗈᓥᏛᏛ㝔་ṑ⸆ಖᏛ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ௦⾲ⓗ࡞⓶ឤᰁཎᅉ⳦࡛࠶ࡿ㯤Ⰽࣈࢻ࢘⌫⳦ ࡣࠊࢺࣆ࣮ᛶ⓶⅖㸦$WRSLFGHUPDWLWLV$'㸧ࡢ ቑᝏࡶ㛵㐃ࡋࠊ㞴ࡢせᅉࡢ୍ࡘ࡛࠶ࡿࠋᮏ ◊✲ࡣࠊ㯤Ⰽࣈࢻ࢘⌫⳦ࡢ $' ⓶ࡢ⳦ࡢ╔࣭ᐃ ╔ᛶࢆࠊࣇࣛࢢࣜࣥḞᦆ࣐࢘ࢫ㔝⏕ᆺ࣐࢘ࢫࢆ ⏝࠸᳨࡚ウࡋࠊ$' ⓶ឤᰁࡍࡿᰴ≉᭷ࡢ⓶ᐃ╔ ᶵᵓࢆゎ᫂ࡍࡿࡇࢆ┠ⓗࡍࡿࠋᮏ◊✲ࡢ㐙⾜ ࡼࡾࠊ$' ቑᝏ࠾ࡅࡿ㯤Ⰽࣈࢻ࢘⌫⳦ࡢึᮇࡢឤ ᰁ࣓࢝ࢽࢬ࣒ࢆ⌮ゎࡋࠊ៏ᛶᝈࡢண㜵ⓗ࡞ࢥࣥࢺ ࣮ࣟࣝࡘ࡞ࡀࡿሗᇶ┙ࢆ☜❧ࡍࡿࡀ࡛ࡁࡿ ⪃࠼ࡽࢀࡿࠋ 㸺ᨭࡢෆᐜ㸼 㯤Ⰽࣈࢻ࢘⌫⳦㸦ࢤࣀ࣒ࢧࢬ⣙ 0E㸧ࡢ $'㸦 ࢺࣆ࣮ᛶ⓶⅖㸧ࡢ㑇ఏⓗᇶ┙ࢆ᫂ࡽࡍࡿࡓࡵ ࠊ⏦ㄳ᭩グ㍕ࡋࡓ㸱ࡘࡢ◊✲ィ⏬ᚲせ࡞ィ㸯 㸶㸰ᰴࡢࢤࣀ࣒ゎㄞࢆ౫㢗ࡋࡓࠋ㸯㸶㸰ᰴࡣ 0L6HT ࡚ゎㄞࡋ࡚㡬࠸ࡓࠋຍ࠼࡚ࠊྛ⣔⤫ࡢ௦⾲㸯㸮ᰴ ࡘ࠸࡚ࡣࠊࢥࣥࣉ࣮ࣜࢺࢤࣀ࣒ࢆྲྀᚓࡍࡿࡓࡵࠊ 3DF%LR ཬࡧ +L6HT ࡛ࡢゎㄞࢆ⾜ࡗ࡚㡬࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㯤Ⰽࣈࢻ࢘⌫⳦ࡢࢤࣀ࣒ゎㄞࡢࡓࡵࡢࣛࣈࣛࣜ ࣮ㄪᩚ࠾ࡅࡿၥ㢟Ⅼࡀ᫂ࡽ࡞ࡗࡓࠋIllumina ♫ࡢ Nextera ࡛ࣛࣈ࣮ࣛࣜసᡂࡋࡓሙྜࠊ㧗࠸ࢩ ࣮ࢣࣥࢫࣂࢫ㸦ࢩ࣮ࢣࣥࢫ࢝ࣂ࣮ࣞࢪࡢᆒ୍ ᛶ㸧ࡀ⏕ࡌࡿࡇࡀㄆࡵࡽࢀࡓࠋࡑࡢⅭࠊKmer 㢖 ᗘࡀపࡃࠊࢭࣥࣈࣝຠ⋡ࡀ㠀ᖖᝏࡗࡓࠋࡑࡇ ࡛ࠊNextera ࡢࡼ࠺࡞ transposome ࡼࡿ DNA ᩿ ∦ adaptor ligation ࢆྠ⾜࠺ᡭἲ࡛ࡣ࡞࠸ ࡢ࢟ࢵࢺࢆ⏝࠸࡚ẚ㍑᳨ウࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊ 㯤Ⰽࣈࢻ࢘⌫⳦࡛ࡣᚑ᮶ἲࡢࣛࣈ࣮ࣛࣜㄪᩚ࢟ࢵ ࢺ㸦Illumina ♫ TruSeq ࡸ KAPA Biosystem ♫࢟ࢵ ࢺ࡞㸧࡛ࡣⰋዲ࡞ Kmer 㢖ᗘࡢࣆ࣮ࢡࡀㄆࡵࡽࢀࠊ ࢩ࣮ࢣࣥࢫࣂࢫࡣపࡗࡓࠋࡋࡓࡀࡗ࡚ࠊ㯤Ⰽ ࣈࢻ࢘⌫⳦࡛ࡢࣛࣈ࣮ࣛࣜㄪᩚἲ࠾ࡅࡿࢩ࣮ࢣ ࣥ ࢫ ࣂ ࢫ ࡢ ⌧ ㇟ ࡣ ࠊ transposome ࢆ ⏝ ࠸ ࡓ Nextera ⣔㝈ࡗ࡚⏕ࡌࡿ⌧㇟࡛࠶ࡿࡇࡀุ᫂ࡋ ࡓࠋࡇࡢணഛ᳨ウࡢ⤖ᯝࢆ㋃ࡲ࠼ࠊ182 ᰴࡢࢻࣛࣇ ࢺࢤࣀ࣒㓄ิࢆᚓࡿࡓࡵࠊࣛࣈ࣮ࣛࣜㄪᩚ࢟ࢵ ࢺࡋ࡚ࠊ ᭱ࡶ high-throughput 㐺ࡋ࡚࠸ࡿ KAPA Biosystem ♫ HyperPlus kit ࢆ᥇⏝ࡋ࡚ MiSeq ࣮ࣜ ࢻࢆྲྀᚓࡋࡓ㸦2016/1/20 㓄ิࢹ࣮ࢱࢆእࡅ HDD ࡚ཷྲྀࡗࡓ㸧 ࠋ ୍᪉ࠊྛ⣔⤫ࡢ௦⾲㸯㸮ᰴࡘ࠸࡚ࡣࠊPacBio ཬ ࡧ HiSeq ࡛ゎㄞࡋ࡚࠸ࡓࡔࡁࠊPacBio ࣮ࣜࢻࡢࡳ࡛ ࢚࣮ࣛಟṇ࠾ࡼࡧࢭࣥࣈࣜࢆ⾜ࡗ࡚㡬࠸ࡓᚋࠊࡑ ࢀ HiSeq ࡢ࣮ࣜࢻࢆ࣐ࢵࣆࣥࢢࡍࡿࡇ᳨࡛ฟࡉ ࢀࡓ࢚࣮ࣛࡶಟṇࡋࡓ⎔≧㓄ิࡀᚓࡽࢀࡓ 㸦2016/1/4 ࢭࣥࣈࣝࢹ࣮ࢱࢆ࣓࣮ࣝῧ࡚ཷ ྲྀࡗࡓ㸧 ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 㓄ิࢹ࣮ࢱࡣᥞࡗࡓࡢ࡛ࠊᚋࡢணᐃࡋ࡚ࠊ⏦ ㄳ᭩グ㍕ࡋࡓ㸱ࡘࡢ◊✲ィ⏬ᚑࡗ࡚ࠊሗゎᯒ ࢆ⾜࠸ࠊAD ⏤᮶㯤Ⰽࣈࢻ࢘⌫⳦ࢡ࣮ࣟࣥࡢࢺࣆ࣮ ᛶ⓶ࡢ⳦ࡢ╔࣭ᐃ╔ᶵᵓࢆศᏊ⏕≀Ꮫⓗ✲ ᫂ࡍࡿࡇࠊຍ࠼࡚ࠊᕷ୰ឤᰁᆺ MRSA-ST8 ࢢ࣮ࣝ ࣉࡢẚ㍑ࢤࣀ࣒ゎᯒࡼࡾࠊ⢾ᛂ⟅ᛶࡢẘ⣲⏘⏕ ไᚚᶵᵓࡢゎᯒࢆ⾜࠺ணᐃ࡛࠶ࡿࠋ ࣛࣈ࣮ࣛࣜ200 ᰴࡢᰴࡢࢤࣀ࣒㓄ิࢹ࣮ࢱࠊ ࡞ࡽࡧࠊྛ⣔⤫ࡢ௦⾲ᰴࡢࢤࣀ࣒㓄ิࡀྲྀᚓ ࡛ࡁࡓࡇ࡛ࠊࡇࢀࡲ࡛ࡢ CGH ࣮࣋ࢫࡢ⣔⤫ẚ㍑ ࡽࠊࢤࣀ࣒㓄ิ࡛ࣞ࣋ࣝࡢࡼࡾ㧗⢭ᗘ࡞⣔⤫ẚ㍑ ゎᯒࡀྍ⬟࡞ࡿࠋࡉࡽࠊẘ⣲㑇ఏᏊࢆᣢࡘࣇ ࣮ࢪࡸ MRSA ᆺࡢ Clade ࡞ࡢ㐍⣔⤫ゎᯒࡶ⾜ࡗ ࡚࠸ࡃணᐃ࡛࠶ࡿࠋ ࡉࡽࠊ㯤Ⰽࣈࢻ࢘⌫⳦ࢤࣀ࣒ࡢࣛࣈ࣮ࣛࣜㄪ ᩚἲࡢ㐪࠸࡛ࠊࢩ࣮ࢣࣥࢫࣂࢫࡀᏑᅾࡍࡿࡇ ࡀุ᫂ࡋࡓࡇࡽࠊఱᨾࡇࡢࡼ࠺࡞ࢩ࣮ࢣࣥࢫࣂ ࢫࡀ⏕ࡌ࡚ࡋࡲ࠺ࡢࠊᚋࠊࡑࡢ࣓࢝ࢽࢬ࣒ ࡢゎᯒࡀᚲせ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 2016/1/20 㓄ิࢹ࣮ࢱࢆཷྲྀࡗࡓẁ㝵࡞ࡢ࡛ࠊࡇ ࢀࡽࠊヲ⣽࡞ࢤࣀ࣒ゎᯒࡀ㐍⾜ࡍࡿࠋࡑࡢࡓࡵࠊ ㄽᩥ➼ࡢබ⾲ࡋࡓ㡯┠ࡣ࠶ࡾࡲࡏࢇࠋ - 387 - බເᨭㄢ㢟㸸2015 ᖺᗘ ື≀ከ⣽⬊ᛶ㐍࣓࢝ࢽࢬ࣒ࡢศᏊ⏕≀Ꮫⓗゎ᫂ ◊✲άືࢫࢱ࣮ࢺᨭ Ⳣ ⿱࣭┴❧ᗈᓥᏛ⏕⎔ቃᏛ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ື≀ࡢከ⣽⬊యไࡢ㐍ࡣࡢࡼ࠺࡞㑇ఏᏊࡀ ᪂ࡓసࡽࢀࡿࡇࡀᚲせࡔࡗࡓࡢ㸽ᡈ࠸ࡣࡢ ࡼ࠺࡞㑇ఏᏊࡀ༢⣽⬊ࡢ௦ࡽᏑᅾࡋࠊࡑࢀࡀከ ⣽⬊యไࡢᵓ⠏࠶ࡓࡾ㌿⏝ࡉࢀࡿࡼ࠺࡞ࡗࡓࡢ 㸽ࡇ࠺ࡋࡓၥ㢟ࡢゎỴࡣࠊື≀⣔⤫ⓗ㏆⦕ ࡞༢⣽⬊⏕≀ࡢࢤࣀ࣒ࢆከᩘゎㄞࡋࠊື≀ࡸࡢཎ ⏕⏕≀ࡢࢤࣀ࣒ẚ㍑ࡍࡿࡇࡀᚲせ࡛࠶ࡿࠋࡋ ࡋ࡞ࡀࡽࠊ⌧ᅾࠊື≀ࡢࢤࣀ࣒ࡣከᩘỴᐃࡉࢀ࡚࠸ ࡿࡶࡢࡢࠊື≀㏆⦕࡞༢⣽⬊⏕≀ࡢࢤࣀ࣒ࡣ㸱✀ 㢮ࡋỴᐃࡉࢀ࡚࠸࡞࠸ࠋಙ㢗ᛶࡢ㧗࠸ẚ㍑ࢤࣀ࣒ ゎᯒࡢࡓࡵࡣࠊࡇࡢᩘࢆቑࡸࡍࡇࡀᚲせ࡛࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ື≀㏆⦕࡞༢⣽⬊⏕≀ࡋ࡚ࡣࠊᚑ᮶❧あ㠴ẟ ࡀ▱ࡽࢀ࡚࠸ࡓࡀࠊ᭱㏆ࠊࢡࢳ࢜ࢫ࣏ࣞ ࡤࢀࡿཎ⏕⏕≀ࢢ࣮ࣝࣉࡀⓎぢࡉࢀࡓࠋࡑࡢ࠺ࡕࡢ 㸰 ✀ ࠊ Amoebidium parasiticum ࠾ ࡼ ࡧ Sphaeroforma arctica ࡢࢤࣀ࣒㓄ิỴᐃࢆ⾜࠺ࠋ ணഛⓗ࡞ゎᯒࡽࠊࡇࡢ㸰✀ࡣࢤࣀ࣒୰ከࡃࡢ⧞ ࡾ㏉ࡋ㓄ิࢆᣢࡕࠊ㓄ิỴᐃࡀ࡞ࡾᅔ㞴࡛࠶ࡿࡇ ࡀศࡗ࡚࠸ࡿࠋࡑࡇ࡛ᨭഃࡢᑓ㛛ⓗ࡞▱㆑ ᢏ⾡ࡢຓࡅࢆࡾࡓ࠸ࠋࡶࡋᚲせ࡛࠶ࢀࡤࠊPacBio ࡢ㉸ࣟࣥࢢ࣮ࣜࢻࡶྵࡵ࡚ᨭࢆ࠾㢪࠸ࡍࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ⌧Ⅼ࡛ࡣࡲࡔࢹ࣮ࢱࡀ࡞ࡃࠊࡋࡓࡀࡗ࡚ゎᯒࡶ ⾜ࢃࢀ࡚࠸࡞࠸ࡀࠊࣉࣟࢪ࢙ࢡࢺᚋࡢᒎ㛤ࡋ ࡚ࡣࠊ௨ୗࡀணࡉࢀࡿ 㸯㸧ẚ㍑ࢤࣀ࣒ゎᯒ ᪤Ꮡࡢື≀ࢤࣀ࣒❧あ㠴ẟࢤࣀ࣒ࠊཬࡧࣇ ࣛࢫࢸࣞࢤࣀ࣒ྜࢃࡏ࡚ࠊẚ㍑ࢤࣀ࣒ࡢ ᡭἲࡼࡿࠕື≀ከ⣽⬊యไ㐍ࡢศᏊ࣓࢝ࢽ ࢬ࣒ࠖ◊✲ࢆᒎ㛤ࡉࡏࡿࠋ≉ࠊື≀ከ⣽⬊ᛶ ࡢ㐍࠶ࡓࡾࡢࡼ࠺࡞㑇ఏᏊࡀቑຍࡋࡓࡢ ࠊࡢࡼ࠺࡞㑇ఏᏊࡀ᪂ࡋࡃసࡽࢀࡓࡢࠊ ࠶ࡿ࠸ࡣ᪂ࡋࡃసࡽࢀࡓࡢ࡛ࡣ࡞ࡃ᫇ࡽᏑᅾ ࡋࡓࡢࠊ╔┠ࡋ࡚ゎᯒࢆ⾜࠺ࠋ 㸰㸧༢⣽⬊⏕≀ࡀᣢࡘࠕከ⣽⬊ⓗ࡞ࠖ㑇ఏᏊࡢᶵ⬟ ゎᯒ ẚ㍑ࢤࣀ࣒ゎᯒࡼࡾⓎぢࡉࢀࡓࠊ୍ぢ༢⣽⬊ ⏕≀ࡣᚲせࡀ࡞࠸ᛮࢃࢀࡿࠕከ⣽⬊ⓗ࡞ࠖ㑇 ఏᏊࡢᶵ⬟ゎᯒࢆ⾜࠺ࠋ≉ࠊᚓࡽࢀࡓࢤࣀ࣒㓄 ิࢆά⏝ࡋࠊࢤࣀ࣒⦅㞟ࡢ㐺⏝㔜Ⅼࢆ⨨ࡃࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 2015 ᖺ㸷᭶㸯㸳᪥ࢧࣥࣉࣝࢆ㏦ࡋࡓࠋ⌧ᅾ㓄 ิỴᐃసᴗ୰࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾࡢࡇࢁ࡞ࡋࠋ - 388 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࣂ࢜ࣇ࣒࣐ࣝࢺࣜࢡࢫᡂศࡢ᪂ᶵ⬟ࡢ㛤ᣅ ⱝᡭ◊✲㸦㸿㸧 ᮡᮏ ┿ஓ࣭ᮾிឿᜨ་⛉Ꮫ་Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࣂ࢜ࣇ࣒ࣝࡣᚤ⏕≀ࡼࡾᙧᡂࡉࢀࡿ⭷≧ࡢᵓ 㐀య࡛࠶ࡾࠊ་⒪ࡢ⌧ሙ࡛ࡣࠊ⾑⟶ෆ␃⨨࢝ࢸ࣮ࢸ ࣝࡸ࣮࣌ࢫ࣓࣮࣮࢝࡞ࡢ་⒪ჾලࡢ⾲㠃ᙧᡂࡉ ࢀࡿࠋࣂ࢜ࣇ࣒ࣝࢆᙧᡂࡍࡿཎ⣽⳦ࡢ࡞࡛ ࡶ㯤Ⰽࣈࢻ࢘⌫⳦ࡣࠊࣄࢺ✀ࠎࡢ⮋ᛶᝈ࣭⫵ ⅖࣭㦵㧊⅖࣭ᚰෆ⭷⅖࡞ࢆᘬࡁ㉳ࡇࡋࠊせ࡞㝔 ෆឤᰁ⳦ࡋ࡚㔜せどࡉࢀ࡚࠸ࡿࠋࣂ࢜ࣇ࣒ࣝ ෆ㒊ࡢ⣽⳦ࡣᢠ⳦ࡸච⣔࣭㈎㣗⣽⬊⣔ࡢാࡁ 㧗ᗘࡢ⪏ᛶࢆ♧ࡍࡓࡵࠊࣂ࢜ࣇ࣒ࣝ㛵㐃ࡍࡿ ឤᰁ㸦ࣂ࢜ࣇ࣒ࣝឤᰁ㸧ࡣ៏ᛶ࣭㞴 ࡢ୍㏵ࢆࡓࡿࡇࡀከ࠸ࠋ⌧௦♫࠾࠸࡚ࠊࣂ ࢜ࣇ࣒ࣝឤᰁࡢⓗ☜࡞ண㜵ἲ࣭⒪ἲࡢ㛤Ⓨ ࡣႚ⥭ࡢㄢ㢟࡛࠶ࡿࠋ ࡇࢀࡲ࡛⏦ㄳ⪅ࡽࡣࠊࣂ࢜ࣇ࣒ࣝឤᰁࡢ せ࡞㉳ᅉ⳦࡛࠶ࡿ㯤Ⰽࣈࢻ࢘⌫⳦ࡢࣂ࢜ࣇࣝ ࣒࣐ࢺࣜࢡࢫࠊపศᏊ 51$ ࡀᏑᅾࡋࠊࣂ࢜ࣇ ࣒ࣝᙧᡂ㔜せ࡛࠶ࡿࡇࢆⓎぢࡋࡓࠋࡋࡋࠊࡇ ࢀࡽࡢపศᏊ 51$ ࡀ⣽⳦ࡼࡗ࡚⏘⏕㸦ศἪ㸧ࡉࢀ ࡓࡶࡢ࡞ࡢࠊࡶࡋࡃࡣᇵᆅ㸦እ⎔ቃ㸧୰ࡢ 51$ ࢆ ᤕᤊࡋࡓࡶࡢ࡞ࡢࡣ࡛᫂࠶ࡗࡓࠋࡑࡇ࡛ᮏ◊✲ ࡛ࡣࠊࣂ࢜ࣇ࣒࣐ࣝࢺࣜࢡࢫ୰ྵࡲࢀࡿపศ Ꮚ 51$ ࠾ࡼࡧᇵᆅ୰ࡢ 51$ ࡢሷᇶ㓄ิࢆ⥙⨶ⓗゎ ᯒࡍࡿࠋࡑࢀ୪⾜ࡋ࡚ࠊࣂ࢜ࣇ࣒ࣝࡢᙧᡂ ࠾࠸࡚㔜せ࡛࠶ࡿࡀࠊࡑࡢ㓄ิࡸᵓ㐀࡞᫂࡞Ⅼ ࡢከ࠸⣽⬊እ '1$ ࡘ࠸࡚ࡶሷᇶ㓄ิࢆỴᐃࡋࠊࢤ ࣀ࣒㓄ิẚ㍑ࡍࡿࠋࡇࢀࡽࡼࡾࠊࣂ࢜ࣇࣇ ࣒ࣝ㔜せ࡞⣽⬊እ᰾㓟ࡢᐇయゎ᫂ࢆ┠ᣦࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࣂ࢜ࣇ࣒ࣝᙧᡂ⬟ࡢ㧗࠸㯤Ⰽࣈࢻ࢘⌫⳦⮫ᗋศ 㞳ᰴ ᰴ㸦05 ᰴཬࡧ 05 ᰴ㸧ࡘ࠸࡚ࡢ᪂つࢤࣀ ࣒㓄ิ⣽⬊እ '1$ 㓄ิ㸦 ᳨య㸧ࢆỴᐃࡋࡓࠋ ࡑࡢᚋࠊᚓࡽࢀࡓࢤࣀ࣒㓄ิୖ⣽⬊እ '1$ ࡢ࣮ࣜ ࢻࢆ࣐ࢵࣆࣥࢢࡋࡓ㸦ࠊᯘ *㸧ࠋࡲࡓࠊ05 ᰴࡢ ࣂ࢜ࣇ࣒࣐ࣝࢺࣜࢡࢫࠊᇵ㣴๓ࡢᇵᆅཬࡧᇵ㣴 ⤊ࡢᇵᆅୖΎྵࡲࢀࡿ 51$ ࢆ 51$VHT 㸦 ᳨య㸧 ࡼࡾゎᯒࡋࡓ㸦ᮾࠊⳢ㔝࣭㕥ᮌ *㸧 ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣂ࢜ࣇ࣒ࣝᙧᡂ⬟ࡀ㧗ࡃࠊࡑࢀࡽࡢᛶ㉁ࡀࡃ ␗࡞ࡿ㯤Ⰽࣈࢻ࢘⌫⳦⮫ᗋศ㞳ᰴ ᰴ㸦⣽⬊እከ⢾ ౫Ꮡࡋࡓࣂ࢜ࣇ࣒ࣝࢆᙧᡂࡍࡿ 05 ᰴཬࡧࢱ ࣥࣃࢡ㉁౫Ꮡࡋࡓࣂ࢜ࣇ࣒ࣝࢆᙧᡂࡍࡿ 05 ᰴ㸧ࡢࢻࣛࣇࢺࢤࣀ࣒㓄ิࢆỴᐃࡋࡓࠋ⌧ᅾࠊࣀ ࢸ࣮ࢩࣙࣥࡢ ᰴ㛫࠾ࡅࡿẚ㍑ࢤࣀ࣒ゎᯒ ࢆ㐍ࡵ࡚࠸ࡿࠋࡲࡓࠊᚓࡽࢀࡓࢻࣛࣇࢺࢤࣀ࣒ᑐ ࡋ࡚ࢤࣀ࣒ '1$ ཬࡧ⣽⬊እ '1$ ࡢ࣮ࣜࢻࢆࡑࢀࡒࢀ ࣐ࢵࣆࣥࢢࡋࡓࡇࢁࠊࡑࢀࡽࡢࣃࢱ࣮ࣥࡸ࢝ࣂ࣮ ⋡ࡣ㓞ఝࡋ࡚࠸ࡓࠋࡇࡢࡇࡽࠊ⣽⬊እ '1$ ࡣ⁐ ⳦➼ࡼࡾࢤࣀ࣒㛗ࡀ⣽⬊እฟࡉࢀࠊࣂ࢜ ࣇ࣒࣐ࣝࢺࣜࢡࢫෆྲྀࡾ㎸ࡲࢀ࡚࠸ࡿྍ⬟ᛶࡀ 㧗࠸⪃࠼ࡽࢀࡿࠋ ࡲࡓࠊࣂ࢜ࣇ࣒࣐ࣝࢺࣜࢡࢫྵࡲࢀࡿ 51$ ࢆゎᯒࡋࡓ⤖ᯝࠊ㯤Ⰽࣈࢻ࢘⌫⳦⏤᮶ࡍࡿ≉ᐃࡢ 51$ ࡀࣂ࢜ࣇ࣒࣐ࣝࢺࣜࢡࢫྵࡲࢀ࡚࠸ࡿࡇ ࡀศࡗࡓࠋࡉࡽࠊࣂ࢜ࣇ࣒ࣝᙧᡂ ⏝ࡋࡓᇵᆅ⏤᮶ࡍࡿ 51$ ࡶࣂ࢜ࣇ࣒ࣝෆྲྀ ࡾ㎸ࡲࢀ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋᚋࠊ㯤Ⰽࣈ ࢻ࢘⌫⳦ࡢࢻࣛࣇࢺࢤࣀ࣒ୖ࣐ࢵࣆࣥࢢࡉࢀ࡞ ࡗࡓ 51$㸦ࡍ࡞ࢃࡕᇵᆅ⏤᮶ࡍࡿ 51$㸧ࡢࣀࢸ࣮ ࢩࣙࣥࡢࢆ⾜࠺ணᐃ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣂ࢜ࣇ࣒ࣝࡢᙧᡂᴟࡵ࡚㔜せ࡞ᙺࢆᯝࡓࡍ ⣽⬊እ '1$ ࡢ⏤᮶ࡸ㓄ิࡣࡇࢀࡲ࡛࡛᫂࠶ࡗࡓࠋ ࣂ࢜ࣇ࣒ࣝෆࡢ୍㒊ࡢ⳦ࡀ⁐⳦➼ࡼࡾ⮬㌟ࡢ ࢤࣀ࣒ '1$ ࢆ⣽⬊እฟࡍࡿྍ⬟ᛶࠊఱࡽࡢ ࣓࢝ࢽࢬ୍࣒࡛㒊ࡢ㓄ิࢆ⬟ືⓗࡘ㑅ᢥⓗศἪ ࡍࡿྍ⬟ᛶࡘ࠸࡚ࠊ㛗ࡽࡃ㆟ㄽࡉࢀ࡚ࡁࡓࠋᮏ◊ ✲࡛ᚓࡽࢀࡓᡂᯝࡣࠊࡇࡢㄽத୍▼ࢆᢞࡌࡿ㔜せ ࡞▱ぢ࡛࠶ࡾࠊᚋ⪅ࢆᨭᣢࡍࡿࡶࡢ࡛࠶ࡿࠋ ࡲࡓࠊ࠶ࡿ✀ࡢ⣽⳦࠾࠸࡚ࠊ51$ ࡀ⣽⬊እᏑᅾ ࡍࡿࡇࡀࡍ࡛ሗ࿌ࡉࢀ࡚࠸ࡓࡀࠊ⣽⬊እ࡛ࡢ 51$ ࡢᶵ⬟ࡸฟ㸦ศἪ㸧ࡢ࣓࢝ࢽࢬ࣒ࡣ༑ศ⌮ゎࡉ ࢀ࡚࠸࡞࠸ࠋᮏ◊✲࡛ࡣࠊ⣽⬊እฟࡉࢀࡓపศ Ꮚ 51$ ࡀࣂ࢜ࣇ࣒ࣝᙧᡂ࠾࠸࡚㔜せ࡞ᙺࢆ ᯝࡓࡍ࠸࠺᪂ࡋ࠸⌧㇟ࡢⓎぢᇶ࡙ࡁࠊࡑࡢᐇែ ゎ᫂ࡢ⣒ཱྀࢆᥗࡴࡇࡀฟ᮶ࡓࠋᮏᨭㄢ㢟ࢆ᥎㐍 ࡍࡿࡇࡼࡾࠊ≉ᐃࡢሷᇶ㓄ิࢆᣢࡗࡓ᰾㓟ࡀࣂ ࢜ࣇ࣒ࣝᙧᡂ㔜せ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿࡇ ࡀ᫂ࡽ࡞ࢀࡤࠊࡑࢀࡽࡢ✵㛫ⓗ࡞ືែ㸦ᒁᅾ ኚ࡞㸧ࡢほᐹࢆྍ⬟ࡍࡿ࢜ࣜࢦࢾࢡࣞ࢜ࢳࢻ ࡸࠊࣂ࢜ࣇ࣒ࣝᙧᡂࢆไᚚࡋ࠺ࡿࣉࢱ࣐࣮ࡢ タィࡶ⧅ࡀࡿ⪃࠼ࡽࢀࡿࠋࡇࢀࡽࡢࢤࣀ࣒⛉Ꮫ ❧⬮ࡋࡓ◊✲ࢆᒎ㛤ࡍࡿࡇ࡛ࠊ51$ ࢆᕠࡿ᪂ࡓ࡞ ᴫᛕࡢㄌ⏕ࣂ࢜ࣇ࣒ࣝឤᰁࡢண㜵ἲ࣭⒪ ἲࡢ㛤Ⓨ⧅ࡀࡿᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 389 - බເᨭㄢ㢟㸸2013 ᖺᗘ ᰿⢏ඹ⏕࠾ࡅࡿᐟ⣽⬊᰿⢏⳦ࡢ┦ㄆ㆑ᶵᵓࡢゎᯒ ᪂Ꮫ⾡㡿ᇦ◊✲ ᑑᓮ ᣅဢ࣭ᇶ♏⏕≀Ꮫ◊✲ᡤඹ⏕ࢩࢫࢸ࣒◊✲㒊㛛 ࠾ࡅࡿᐟ࣭᰿⢏⳦᪉ࡢ㑇ఏᏊⓎ⌧ࣉࣟࣇ 㸺⫼ᬒ࣭┠ⓗ㸼 ࣐࣓⛉᳜≀᰿⢏⳦ࡢඹ⏕ჾᐁ࡛࠶ࡿ᰿⢏ࡢึ ࣝࢆྲྀᚓࡍࡿࡓࡵࡢࠊ᰿⢏⳦ឤᰁึᮇࡢ㔝⏕ᆺ ᮇⓎ⏕㐣⛬࡛ࡣࠊ᰿⢏⳦ឤᰁࡼࡗ࡚ᐟ᳜≀᰿ࡢ YDJ ኚ␗యࡑࢀࡒࢀ࠾ࡅࡿ 51$VHT ゎᯒࡢᨭࢆ ୍㒊ࡢ⓶ᒙ⣽⬊ࡀ⬺ศࡋࠊ⣽⬊ศࡀㄏᑟࡉࢀ᰿ ཷࡅࡓࠋ ⢏ཎᇶࡀᙧᡂࡉࢀࡿࠋࡑࡢ୍᪉࡛ࠊ᰿⢏⳦ࡣឤᰁ⣒ ࡤࢀࡿ⟶≧ࡢᵓ㐀ࢆࡋ࡚ᐟ⣽⬊ධࡍ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡿࡇࡀ࡛ࡁࡿࠋ᰿⢏ඹ⏕ࡢᡂ❧ࡣࠊࠕ᰿⢏ࡢⓎ ᐟ᳜≀ഃࡢ 51$VHT ࢆ⾜࠸ࠊYDJ ኚ␗య࠾࠸࡚ ⏕ࠖࠕឤᰁ⣒ࢆࡋࡓ᰿⢏⳦ឤᰁࠖࡀྠㄪⓗ㐍 ≉␗ⓗⓎ⌧ኚືࡍࡿ㑇ఏᏊ⩌ࢆྠᐃࡋࡓࠋࡲࡓࠊ ⾜ࡍࡿࡇࡀᚲせྍḞ࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚࠸ ឤᰁ⣒୰Ꮡᅾࡍࡿ᰿⢏⳦ഃࡢ㑇ఏᏊࡘ࠸࡚ࡣࠊ ࡿࡀࠊಶࠎࡢ㇟ࡢไᚚᶵᵓࡸࠊࡑࡢྠㄪᛶࢆไᚚ 51$VHT ࡼࡾࠊ᰿⢏⳦ឤᰁⓎ⌧ㄏᑟࡉࢀࡿ ࡍࡿᶵᵓࡣᮍゎ᫂࡞Ⅼࡀከ࠸ࠋᮏ◊✲ࡣࠊ᰿⢏Ⓨ⏕ 㑇ఏᏊࢆྠᐃࡋࡓࠋࡇࡢ࠺ࡕࠊ 㑇ఏᏊࡣ YDJ ኚ␗ ࠾ࡼࡧឤᰁ⣒ᙧᡂࡢ㑇ఏⓗไᚚᶵᵓࢆ᫂ࡽࡍ య࠾࠸࡚Ⓨ⌧ㄏᑟࡀ㉳ࡇࡽ࡞࠸ࡇࡀࢃࡗࡓࠋ ࡿࡇࡼࡾࠊ᰿⢏ඹ⏕ᡂ❧ࡢᇶ┙࡞ࡿᐟ⣽⬊ ᰿⢏⳦ࡢ㛫࡛⾜ࢃࢀࡿሗࡢࡸࡾࡾࡢศᏊ࣓ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢝ࢽࢬ࣒ࢆゎ᫂ࡍࡿࡇࢆ┠ⓗࡋ࡚࠸ࡿࠋ ឤᰁ⣒ࡢ୰Ꮡᅾࡍࡿ᰿⢏⳦ࡣࡁࢃࡵ࡚ᑡᩘ࡛࠶ ࡿࡓࡵࠊࡑࢀ⏤᮶ࡍࡿ᰿⢏⳦ 51$ ࡶᴟᚤ㔞࡛࠶ࡗ 㸺ᨭࡢෆᐜ㸼 ࡓࡀࠊ60$57HU ἲࡼࡾࠊᴟᚤ㔞 51$ ࡽ F'1$ ࣛ ࣐ ࣓ ⛉ ࡢ ࣔ ࢹ ࣝ ᳜ ≀ ࣑ ࣖ ࢥ ࢢ ࢧ ࡢ YDJUDQW ࣈ࣮ࣛࣜࢆసᡂࡍࡿࡇᡂຌࡋࡓࠋឤᰁ⣒୰ࡢ᰿ LQIHFWLRQWKUHDGYDJኚ␗య࡛ࡣࠊ᰿⢏ࡢᙧ ⢏⳦ࡢ 51$VHT ゎᯒࡣࡇࢀࡲ࡛ሗ࿌ࡀ࡞ࡃࠊᮏ◊✲ ᡂࡀ࡞ࡿ୍᪉࡛ࠊឤᰁ⣒ࡢᙧᡂࡣ୍ぢṇᖖ ࡢ㐍ᒎࡣ᰿⢏ඹ⏕ࡢศᏊ࣓࢝ࢽࢬ࣒ࡢ⌮ゎ㈉⊩ࡍ ㉳ࡇࡿࠋࡋࡋࠊಶࠎࡢឤᰁ⣒ࡣ⾲⓶ୖࢆࡉࡲࡼ࠺ ࡿࡔࡅ࡛࡞ࡃࠊ᳜≀ᚤ⏕≀┦స⏝◊✲ࡢ᪂ࡓ࡞᪉ ࡼ࠺ఙ㛗ࡋࠊ⓶ᒙ⣽⬊ධࡍࡿࡇࡀ࡛ࡁ࡞࠸ࠋ ἲㄽࢆᥦ♧࡛ࡁࡿ⪃࠼࡚࠸ࡿࠋ ࡇࡢࡇࡽࠊYDJ ኚ␗య࡛ࡣࠊ᰿⢏Ⓨ⏕ࣉࣟࢢࣛ ࣒ࡢ␗ᖖࡔࡅ࡛࡞ࡃࠊ᰿⢏⳦ࢆᐟ⣽⬊ㄏᑟࡍ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡿᶵᵓࡶ◚⥢ࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡿࠋYDJ ኚ␗యࡢ ࡞ࡋ ≉ᚩⓗ࡞ឤᰁ⣒ࡢ⾲⌧ᆺࡽࠊYDJ ኚ␗ࡣ᰿⢏⳦ࡢ ᐟ⣽⬊ࡢㄏᑟ㛵ࢃࡿᐟ᳜≀ഃࡢ㑇ఏᏊࡢࡳ ࡞ࡽࡎࠊᐟࡽࡢࢩࢢࢼࣝࡢᛂ⟅㛵ࡍࡿ᰿⢏ ⳦ഃࡢ㑇ఏᏊࡢⓎ⌧ࣃࢱ࣮ࣥࡶᙳ㡪ࢆཬࡰࡋ࡚࠸ ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋࡑࡇ࡛ࠊ᰿⢏ึᮇⓎ⏕㐣⛬ - 390 - බເᨭㄢ㢟㸸2012 ᖺᗘ ⮬ືᐇ㦂㐍ࢩࢫࢸ࣒ࡼࡿከ⪏ᛶ⳦ࡢฟ㐍ࣉࣟࢭࢫࡢゎᯒ ⱝᡭ◊✲㸦㹀㸧 㕥ᮌ ┿࿃࣭⌮Ꮫ◊✲ᡤ⏕ࢩࢫࢸ࣒◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ⏕≀ࡢᰂ㌾ࡘ㡹ᙉ࡞㐍⬟ࢆ⌮ゎࡋไᚚࡍࡿࡇ ࢆ┠ᣦࡋࠊᮏㄢ㢟࡛ࡣࡁ࡞♫ၥ㢟࡞ࡗ࡚࠸ ࡿከ⪏ᛶ⳦ࢆᑐ㇟ࠊࡑࡢ㐍ࣉࣟࢭࢫࢆゎ᫂ࡍ ࡿࡇࢆලయⓗ࡞┠ⓗࡋࡓࠋ⸆⪏ᛶ⳦ࡢ◊✲ࢆ ᥎㐍ࡋ࡚࠸ࡃୖ୍࡛⯡ⓗ࡞ᡭἲࡣࠊ⮫ᗋ⌧ሙࡽᚓ ࡽࢀࡓ⪏ᛶ⳦ࡢ⾲⌧ᆺ࣭㑇ఏᆺࢆゎᯒࡋ࡚࠸ࡃ࠸ ࠺ࡶࡢ࡛࠶ࡿࠋࡋࡋ࡞ࡀࡽࡇࡢࡼ࠺࡞ᡭἲ࡛ࡣࠊ ⪏ᛶࣉࣟࢭࢫ࠾ࡅࡿ⤒ⓗ࡞ኚࢆ㏣㊧ࡍࡿࡇ ࡀ࡛ࡁ࡞࠸ࠋࡑࡇ࡛ᮏ◊✲࡛ࡣࠊ⮫ᗋ⌧ሙࡽᚓ ࡽࢀࡿ⸆⪏ᛶ⳦ࡇࡔࢃࡽࡎࠊ⸆⪏ᛶ⬟ࢆࡶ ࡓ࡞࠸⭠⳦㔝⏕ᰴࢆᑐ㇟ᐇ㦂㐍ⓗᡭἲࢆ⏝࠸ ࡿࡇ࡛」ᩘ⣔ิࡢ⪏ᛶ⳦ࢆฟࡋࠊࡑࡢ㑇ఏᆺゎ ᯒࢆ⾜࠺ࡇ࡛ከ⪏ᛶࡢ㐍ࣉࣟࢭࢫࡢゎ᫂ࢆ ྲྀࡾ⾜ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 ⪏ᛶ⬟ࢆᣢࡓ࡞࠸⭠⳦㔝⏕ᰴࡽከ⪏ᛶ⳦ࢆ ᐇ㦂㐍ⓗฟࡋࠊࡢࡼ࠺࡞ኚ␗ࡢ✚࠾ࡼࡧ ࡑࢀࡼࡿᶵ⬟ᨵኚࡼࡾከ⪏ᛶࡋᚓࡓࡢ᫂ ☜グ㏙ࡍࡿࡓࡵࠊస⏝ᶵᗎࡢ␗࡞ࡿ 33 ✀㢮ࡢᢠ⏕ ≀㉁ࢆಶูῧຍࡋࡓ⎔ቃୗ࡛⊂❧࡞ 4 ⣔ิ࡛ࡢ㐍 ᐇ㦂ࢆ⾜ࡗࡓࠋࡑࢀࡒࢀࡢ⸆ࡘ࠸࡚ࠊࡑࡢ⃰ ᗘࢆኚࡉࡏࡓ」ᩘࡢ⎔ቃୗ࡛⳦యࢆᇵ㣴ࡋࠊ⳦య ࡢቑṪࡀ☜ㄆࡉࢀࡓ᭱ࡶ㧗࠸⸆⃰ᗘࡢ⎔ቃࡽ 24 㛫࿘ᮇ࡛ḟࡢ⎔ቃ᳜࠼⥅࠸ࡔࠋ࠾ࡼࡑ 3 ࣨ᭶㛫 ࡇࡢ᳜࠼⥅ࡂᇵ㣴ࢆ⾜ࡗࡓࡇࢁࠊ31 ✀㢮ࡢ⸆ ࡘ࠸࡚ቑṪࢆྍ⬟ࡍࡿ⸆⃰ᗘࡢ᭷ព࡞ୖ᪼ࡀぢ ࡽࢀ⸆⪏ᛶ⭠⳦ࡢྲྀᚓᡂຌࡋࡓࠋࡘ࡙࠸࡚ࠊ ࡢࡼ࠺࡞ኚ␗ࡀᑟධࡉࢀࡿࡇࡼࡗ࡚⸆⪏ᛶ ࡀ࡞ࡉࢀࡓࡢゎ᫂ࡍࡿࡓࡵࠊ11 ✀㢮ࡢ⸆ ᑐࡋ࡚ᚓࡽࢀࡓ 44 ᰴࡢ⸆⪏ᛶ⭠⳦ᰴࡢࢤࣀ࣒ ኚ␗ゎᯒࢆḟୡ௦ࢩ࣮ࢣࣥࢧ࡚⾜࡞ࡗࡓࡇࢁࠊ ࡑࢀࡒࢀ 5㹼20 ಶ⛬ᗘࡢሷᇶ⨨࣭ᤄධ࣭Ḟᦆࢆྠ ᐃࡋࡓࠋ ࡇࢀࡽࡢ⤖ᯝࢆඖࠊᮏᨭ࡚㑇ఏᏊኚ␗ࡀ ࡢࡼ࠺࡞㡰␒࡚ᑟධࡉࢀࡓࡢࡑࡢ⤒ኚࡢゎ ᯒࢆ⾜ࡗࡓࠋලయⓗࡣࠊୖグ࡚ྠᐃࡉࢀࡓ㑇ఏ Ꮚኚ␗ࡀ⸆⪏ᛶࣉࣟࢭࢫࡢࡢⅬ࡛ᑟධࡉࢀ ࡓࡢ⤒ⓗ࡞ゎᯒࢆ⾜࠺ࡓࡵࠊ1 㐌㛫ࡈ㸦ィ 12 ᅇ㸧෭ࢫࢺࢵࢡࡉࢀࡓ㐍㏵୰ᰴࡢྠ⟠ᡤࡢሷ ᇶ㓄ิࡢゎᯒࢆ౫㢗ࡋࡓࠋࡇࡢࡼ࠺࡞◊✲࡛ࡣࠊኚ ␗⟠ᡤࡢ PCR ࣥࣉࣜࢥࣥࢆࢧ࣮ࣥ࢞ἲ࡚㓄ิ ゎᯒࡍࡿࡇࡀ୍⯡ⓗ࡞ᡭἲ࡛࠶ࡿࡀࠊᮏ◊✲࡛ࡣ ྜィ࡛࠾ࡼࡑ 5000 ⟠ᡤࡢ PCR ࣥࣉࣜࢥࣥࡢゎᯒ ࡀᚲせ࡞ࡾࠊࡇࢀࢆࡍ࡚ࢧ࣮ࣥ࢞ἲ࡚ゎᯒࡍ ࡿࡇࡣ㠀ᖖᅔ㞴࡛࠶ࡿࠋࡑࡇ࡛ᮏᨭゎᯒ࡛ࡣࠊ ḟୡ௦ࢩ࣮ࢣࣥࢧ㸦FLX junior㸧ࢆ⏝࠸࡚㑇ఏᏊኚ ␗ᑟධࡢࢲࢼ࣑ࢡࢫゎᯒࢆໟᣓⓗ⾜࠺ࡇࡋ ࡓࠋ⸆⪏ᛶ⭠⳦ࡢ᭱⤊㐍ᰴ࡚ྠᐃࡉࢀࡓኚ ␗⟠ᡤࢆࠊࡑࢀࡒࢀࡢ㐍㏵୰ᰴ㸦Ѹ80Υࢫࢺࢵࢡ㸧 ࢆ㗪ᆺ PCR ࡼࡾቑᖜࡋࡓࠋࡇࡢ㝿ࠊ」ᩘᰴ࡚ ྠᵝࡢ㑇ఏᏊኚ␗ࡀᏑᅾࡍࡿሙྜࡣࡑࢀࡒࢀᅛ᭷ ࡢࣂ࣮ࢥ࣮ࢻࢱࢢࢆ PCR ࣉ࣐࣮ࣛຍࡋࠊࡢ ⸆⪏ᛶ㐍⣔ิ⏤᮶ࡍࡿ PCR ࣥࣉࣜࢥࣥ࡞ ࡢ༊ู࡛ࡁࡿࡼ࠺ࡋࡓࠋࡘ࡙࠸࡚ྠࡌ᪥ࢫࢺ ࢵࢡࡉࢀࡓ㐍㏵୰ᰴ⏤᮶ࡢ PCR ࣥࣉࣜࢥࣥࢆ ࡍ࡚ΰྜࡋࣂ࣮ࢥ࣮ࢻࢱࢢࢆຍࡍࡿࡇ࡛ࠊࡑ ࡢ PCR ࣥࣉࣜࢥࣥࡢ⏤᮶ࡀ࠸ࡘࢫࢺࢵࢡࡉࢀࡓ ࡶࡢࢃࡿࡼ࠺ࡋࡓࠋࡇࡢࡼ࠺స〇ࡋࡓ࠾ࡼ ࡑ 5000 ⟠ᡤࡢ PCR ࣥࣉࣜࢥࣥࢆࡍ࡚ΰྜࡋ࡚ FLX junior ࡚㓄ิゎᯒࢆ⾜ࡗࡓᚋࠊࡑࢀࡒࢀࡢࢱ ࢢ㓄ิࢆཧ↷ࡋྠᐃࡉࢀࡓ㑇ఏᏊኚ␗ࡀ⸆⪏ᛶ ࣉࣟࢭࢫࡢࡢⅬ࡛ᑟධࡉࢀࡓࡢゎᯒࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏᨭࡼࡾ㑇ఏᏊኚ␗ࡢᑟධࡉࢀࡓ㡰␒ࡀุ᫂ ࡋࡓࠋࡇࡢ⤖ᯝࢆඖࠊ㑇ఏᏊኚ␗ࢆᐇ㦂㐍ࡉࡏ ࡚࠸࡞࠸ඖᰴྠᐃࡉࢀࡓ㡰␒㏻ࡾᑟධࡋࡓኚ␗ యࣛࣈ࣮ࣛࣜࢆస〇ࡋࠊኚ␗యࣛࣈ࣮ࣛࣜᑐ ࡍࡿྛ⸆ࡢ᭱ᑠቑṪ㜼Ṇ⃰ᗘࢆồࡵࡿࡇ࡛ࠊࡑ ࢀࡒࢀࡢኚ␗ࡢ⪏ᛶ⬟ࡢᐤࢆᐃ㔞ࡋ⪏ᛶ⬟ࡢྥ ୖࢆホ౯ࡍࡿࡇࡀྍ⬟࡞ࡗࡓࠋࡉࡽࠊྠᐃࡉ ࢀࡓኚ␗ࢆಶู⤌ࡳ㎸ࢇࡔࣛࣈ࣮ࣛࣜࡢస〇࠾ ࡼࡧࡑࡢᐤࡶᐃ㔞ࡍࡿࡇ࡛༢⊂ኚ␗ࡼࡿ⪏ᛶ ⬟ࡢྥୖ㔞ࡢྜィ㡰␒㏻ࡾ⤌ࡳ㎸ࢇࡔ㝿ࡢྥୖ 㔞ࢆẚ㍑ࡍࡿࡇࡀྍ⬟࡞ࡗࡓࡢ࡛ࠊᚋࠊ࢚ࣆ ࢫࢱࢩࢫࡢ᭷↓ࡸኚ␗ࡢᐤࡢᗘྜ࠸ࢆ⸆ࡢ✀㢮 ࡸ」ᩘࡢ㐍⣔ิ㛫࠾࠸࡚ẚ㍑ࡍࡿࡇ࡛ࠊ⸆ ⪏ᛶ㐍ࣉࣟࢭࢫࡢ≉ᛶࢆᢳฟࡍࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ㏆ᖺࠊ༢⊂ࡢ⸆ᑐࡋ࡚ࡑࡢ⪏ᛶ⬟ࢆ㐍ࡉࡏ ࡓ⣽⳦㢮ࡣࠊࡢస⏝ᶵᗎࢆᣢࡘ⸆ࡶᕪ⪏ᛶ ࢆ♧ࡍࡇࡸࠊࡑࡢ㏫ࠊ࠶ࡿ⸆ᑐࡋ࡚ࡣඖࡢ ᰴࡼࡾࡶᙅࡃ࡞ࡗ࡚ࡋࡲ࠺࠸࠺㉸ឤཷᛶࢆ♧ࡍࡇ ࡀሗ࿌ࡉࢀ࡚࠾ࡾࠊᡃࠎࡶྠᵝࡢ⤖ᯝࢆほᐹࡋሗ ࿌ࡋ࡚࠸ࡿࠋᕪ⪏ᛶࡸ㉸ឤཷᛶ㛵ࢃࡿ࠸ࡃࡘ ࡢᶵᵓࡀゎ᫂ࡉࢀ࡚࠸ࡿࡀࠊ⌧≧࡛ࡣ᫂࡞Ⅼࡶ ከࠎṧࡉࢀ࡚࠸ࡿࠋᮏᨭࡼࡾᡃࠎࡀᚓࡓኚ␗㑇 ఏᏊࡢᑟධࡉࢀࡓ㡰␒ᵝࠎ࡞⸆ᑐࡍࡿ⪏ᛶ⬟ ࢆẚ㍑ࡍࡿࡇࡼࡾࠊ⪏ᛶᰴࡢࢤࣀ࣒ኚ␗ゎᯒࡔ ࡅ࡛ࡣᚓࡽࢀ㞴࠸⾲⌧ᆺ̾㑇ఏᆺࡢ┦㛵ࢆゎᯒࡍࡿ ࡇࡀྍ⬟࡞ࡗࡓࠋࡇࢀࡽࡢ▱ぢࡣࠊ」㞧࡞ᕪ ⪏ᛶ࠾ࡼࡧ㉸ឤཷᛶࢆᘬࡁ㉳ࡇࡍศᏊᶵᵓࡢゎ᫂ ࡘ࡞ࡀࡿࡶࠊ⸆⪏ᛶ⳦ࡢฟ⌧ࢆᢚไࡍࡿ⸆ ᢞᣦ㔪ࡢタィࡸࠊ᪂ࡓ࡞⸆ࡢ㛤Ⓨࡶࡘ࡞ࡀ ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋࠋ - 391 - බເᨭㄢ㢟㸸2015 ᖺᗘ RNA ࢚ࣆࢪ࢙ࢿࢸࢵࢡࢫ㧗ḟ⏕⌧㇟ ᇶ┙◊✲㸦㹑㸧 㕥ᮌ ຮ࣭ᮾிᏛᏛ㝔ᕤᏛ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⏕యෆ䛾 RNA 䛿㌿ᚋ䛻䝇䝥䝷䜲䝅䞁䜾䜔䜶䝕䜱䝔 䜱䞁䜾䚸RNA ಟ㣭䛸䛔䛳䛯ᵝ䚻䛺䝥䝻䝉䝅䞁䜾䜢ཷ䛡 䛶ᡂ⇍䛩䜛䚹䛣䜜䜎䛷䚸RNA ಟ㣭䛾◊✲䛿 tRNA 䜔 rRNA 䛺䛹䛾Ꮡᅾ㔞䛾ከ䛔 RNA 䜢୰ᚰ䛻ゎᯒ䛜㐍 䜑䜙䜜䛶䛝䛯䛜䚸᭱㏆䛾ḟୡ௦䝅䞊䜿䞁䝃䞊䜢⏝䛧 䛯⥙⨶ⓗ䛺ゎᯒ䛻䜘䜚䚸mRNA 䜔 non-coding RNA ୰ 䛻ᵝ䚻䛺ಟ㣭䛜ぢ䛴䛛䜚䛴䛴䛒䜚䚸䜶䝢䝖䝷䞁䝇䜽䝸䝥 䝖䞊䝮䛸䛔䛖᪂䛧䛔ᴫᛕ䛜ᥦၐ䛥䜜䛶䛔䜛䚹ᮏ◊✲䛷 䛿ḟୡ௦䝅䞊䜿䞁䝇䜢ά⏝䛩䜛䛣䛸䛷䚸RNA ಟ㣭㒊 䛾≉ᐃ䜔 RNA ಟ㣭䛾ᶵ⬟䜢᫂䜙䛛䛻䛩䜛䛣䛸䜢┠ ᣦ䛩䚹ලయⓗ䛻䛿䚸௨ୗ䛾 3 䛴䛾䝃䝤䝔䞊䝬䜢ணᐃ 䛧䛶䛔䜛䚹ձ䝠䝖䛻䛚䛡䜛 A-to-I RNA 䜶䝕䜱䝔䜱䞁䜾䛾 ㄪ⠇ᶵᵓ䚸ղtRNA 䝯䝏䝹㓝⣲Ḟᦆ䛻㉳ᅉ䛩䜛㠀 ೃᛶ X 㐃㙐▱ⓗ㞀ᐖ䛾Ⓨᶵᵓ䚸ճ⏕Ꮫⓗᡭ ἲ䛸ḟୡ௦䝅䞊䜿䞁䝇䜢⤌䜏ྜ䜟䛫䛯 N4 䜰䝉䝏䝹䝅 䝏䝆䞁䛾⥙⨶ⓗྠᐃ䚹 ㏆ᖺࠊRNA ࡢಟ㣭㒊ࢆࢤࣀ࣒࣡ࢻྠᐃࡍࡿ ࡓࡵࠊ⏕Ꮫᡭἲḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮ࢆ⤌ࡳ ྜࢃࡏࡓᵝࠎ࡞ᢏ⾡ࡀ㛤Ⓨࡉࢀࠊ┿᰾⏕≀ࡢ mRNA ࡸ non-coding RNA ࣀࢩࣥ(I)ࡸ N6-࣓ࢳࣝࢹ ࣀࢩࣥ(m6A)࡞ࡢಟ㣭ሷᇶࡀ㔞ྠᐃࡉࢀ࡚࠸ ࡿࠋࡇࢀࡽࡢ㌿ᚋಟ㣭ࡣ⣽⬊ෆ࡛ࡢ㑇ఏᏊⓎ⌧ࢆ ไᚚࡋࠊⓎ⏕ࡸศࠊࢫࢺࣞࢫᛂ⟅࡞ᵝࠎ࡞㔜せ ࡞⏕⌧㇟ࡢ㛵ࡀ᫂ࡽࡉࢀ࡚࠾ࡾࠊRNA ಟ 㣭ࡢ㔜せᛶࡀ㧗ࡲࡾࡘࡘ࠶ࡿࠋ I ಟ㣭ࡣࠊ⣽⬊ࡢ᰾ෆ࡛ᙧᡂࡉࢀࡓᮏ㙐 RNA ࡀ ADAR ࡤࢀࡿᮏ㙐 RNA ≉␗ⓗࢹࣀࢩࣥࢹ ࣑ࢿ࣮ࢫㄆ㆑ࡉࢀࠊࢹࣀࢩࣥ(A)ࡀ⬺࣑ࣀ ࡉࢀ࡚ I ኚࡍࡿࡇ࡛⏕ࡌࡿࠋI ࡣᏛᵓ㐀ୖࠊ ࢢࣀࢩࣥ(G)㢮ఝࡋ࡚࠸ࡿࡓࡵࢩࢺࢩࣥሷᇶ ᑐᙧᡂࡀྍ⬟࡛࠶ࡿࡇࡽሷᇶሗୖࠊA ࡽ G ࡢኚࢆពࡋࠊࡑࢀࡺ࠼ A-to-I RNA ࢚ࢹࢸ ࣥࢢࡤࢀ࡚࠸ࡿࠋI ಟ㣭ࡣ㟋㛗㢮ࡢࢺࣛࣥࢫࢡࣜ ࣉࢺ࣮࣒ෆ㇏ᐩᏑᅾࡍࡿࡇࡀ▱ࡽࢀࠊᙜ◊✲ ᐊ࡛ࡣ⊂⮬ࡢゎᯒᢏ⾡ࢆ⏝࠸ࡿࡇ࡛ࠊࣄࢺᡂே⬻ RNA ୰⣙ 30,000 ⟠ᡤࡢ᪂つ I ಟ㣭㒊ࢆྠᐃࡋ ࡓࠋ ᙜ◊✲ᐊࡢඛ⾜◊✲࡛ ADAR1 ┦స⏝ࡍࡿ ࡘࡢ RNA ࣊ࣜࢣ࣮ࢫ(RNA ࣊ࣜࢣ࣮ࢫ 1, 2 ࡪ) ࢆࣀࢵࢡࢲ࢘ࣥࡋࡓࡇࢁ 3’㠀⩻ヂ㡿ᇦ(UTR)ෆࡢ Alu 㓄ิ୰Ꮡᅾࡍࡿ I ಟ㣭ࡢຠ⋡ࡀ㢧ⴭኚືࡍ ࡿ⌧㇟ࢆぢฟࡋࡓࠋࡇࡢ⤖ᯝࡣ RNA ࣊ࣜࢣ࣮ࢫࡀ RNA ࡢḟᵓ㐀ࢆኚࡉࡏࡿࡇ࡛ RNA ࢚ࢹࢸ ࣥࢢࢆไᚚࡍࡿྍ⬟ᛶࢆ♧၀ࡋ࡚࠸ࡿࠋᮏᨭࡢ ୗ࡛ deep-seq ࢆᐇࡋࠊRNA ࣊ࣜࢣ࣮ࢫࡀไᚚࡍ ࡿ RNA ࢚ࢹࢸࣥࢢ㒊ࡢ⥙⨶ⓗ᥈⣴ࡑࡢᶵ ⬟ゎᯒࢆ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 8 ᮲௳ࡢࣄࢺ⚄⤒⭺ⱆ⭘⣽⬊ A172 (Mock, Luc siRNA, ADAR1 siRNA 2 ✀㢮, RNA ࣊ࣜࢣ࣮ࢫ 1 siRNA 2 ✀㢮, RNA ࣊ࣜࢣ࣮ࢫ 2 siRNA 2 ✀㢮)ࡽ ⢭ 〇 ࡋ ࡓ polyA+ RNA ࡽ Illumina ♫ ࡢ strand-specific ࡞ RNA-seq ࡢ࢟ࢵࢺࢆ⏝࠸࡚ cDNA ࣛࣈ࣮ࣛࣜࢆㄪ〇ࡋࡓᚋࠊᅜ❧㑇ఏ◊࣭ẚ㍑ࢤࣀ ࣒ゎᯒ◊✲ᐊࡢ㇏⏣ᩔඛ⏕㏦ࡋࠊHiseq2500 ࢆ ⏝ࡋ࡚ 150bp ࢚࣌ࣥࢻࡢ Rapid ࣮ࣛࣥࣔࢻ࡛ࠊ 3 ࣇ࣮ࣟࢭࣝศࡢࢩ࣮ࢡ࢚ࣥࢫゎᯒࢆᐇࡋ࡚㡬࠸ ࡓࠋࢩ࣮ࢡ࢚ࣥࢫゎᯒᚋࡢሗゎᯒࡣࠊ◊✲ဨࢆྠ ◊✲ᐊὴ㐵ࡋࠊ㔝ཱྀⱥᶞඛ⏕ࡢࡈᣦᑟࡢୗࠊࣄࢺ ࣜࣇࣞࣥࢫࢤࣀ࣒(hg38)ࡢ࣐ࢵࣆࣥࢢࢆ⾜࠸ࠊ RNA ࢚ࢹࢸࣥࢢຠ⋡ࡢኚືゎᯒࡸ ADAR1 ࣀࢵ ࢡࢲ࢘ࣥࢫࣉࣛࢩࣥࢢ㢖ᗘࡀቑຍࡋࡓ Alu ࢚ ࢟ࢯࣥ㡿ᇦࡢᢳฟࢆ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᚓࡽࢀࡓ࣮ࣜࢻࢆࣄࢺࢤࣀ࣒࣐ࢵࣆࣥࢢࡋࡓ⤖ ᯝࠊྛࢧࣥࣉࣝࡶ⣙ 90%ࡢ࣮ࣜࢻࡀṇࡋ࠸ strand ࡢ᪉ྥ࡛ࢤࣀ࣒࣓ࣛࣥࢺࡉࢀࡓࠋࣜ࣎ࢯ࣮࣒ RNA ࡢΰධ⋡ࡣ 1%௨ୗ࡛࠶ࡾࠊ㠀ᖖⰋዲ࡞⤖ᯝ ࡀᚓࡽࢀࡓࠋ࣐ࢵࣆࣥࢢࢹ࣮ࢱࡽ 30,000 ⟠ᡤࡢ I ಟ㣭㒊ࢆᑐ㇟࢝ࣂࣞࢵࢪࡀ 20 ௨ୖࡢ㒊ࢆᢳ ฟࡋࠊI ಟ㣭ຠ⋡ࢆ⟬ฟࡋࡓࠋRNA ࣊ࣜࢣ࣮ࢫࢆࣀ ࢵࢡࢲ࢘ࣥࡋࡓࢧࣥࣉࣝࢥࣥࢺ࣮ࣟࣝࡢ I ಟ㣭ຠ ⋡ࢆẚ㍑ࡍࡿࡇ࡛ࠊI ಟ㣭ࡀ᭷ពኚືࡋ࡚࠸ࡿ㒊 ࢆከᩘྠᐃࡋࡓࠋࡇࢀࡽࡢ⤖ᯝࡣ 2 ࡘࡢ RNA ࣊ࣜ ࢣ࣮ࢫࡀࣄࢺࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒యࡢ I ಟ㣭 ࢆไᚚࡍࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋᚋࡣࠊኚືࡋࡓ I ಟ㣭㒊࠾ࡅࡿ㓄ิࡸḟᵓ㐀ࡢ≉ᚩࢆ᫂ࡽ ࡋ࡚࠸ࡃணᐃ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭ◊✲ࡼࡾ RNA ࣊ࣜࢣ࣮ࢫࡀࢺࣛࣥࢫࢡ ࣜࣉࢺ࣮࣒యࡢ I ಟ㣭ࢆไᚚࡍࡿྍ⬟ᛶࢆぢฟࡋ ࡓࠋRNA ࢚ࢹࢸࣥࢢࡣࣥࢺࣟࣥࡸ 3’UTR ࡞ ࡢ mRNA ࡢࣉࣟࢭࢩࣥࢢࡸ㑇ఏᏊⓎ⌧ࢆไᚚࡍ ࡿࢩࢫ࢚࣓ࣞࣥࢺࡀᏑᅾࡍࡿ㡿ᇦᏑᅾࡍࡿࡓࡵࠊ RNA ࣊ࣜࢣ࣮ࢫࡼࡿ RNA ࢚ࢹࢸࣥࢢไᚚࢆ ࡋࡓ᪂つࡢ㑇ఏᏊⓎ⌧ᶵᵓࡀᏑᅾࡍࡿྍ⬟ᛶࡀ࠶ ࡿࠋᚋࡣࠊRNA ࣊ࣜࢣ࣮ࢫࡀࡢࡼ࠺࡞࣓࢝ࢽࢬ ࣒ࡀ I ಟ㣭ࡢኚືࢆᘬࡁ㉳ࡇࡍࡸࠊI ಟ㣭ࡢኚືࡀ ලయⓗࡢࡼ࠺࡞⏕⌧㇟㛵ࡍࡿࢆ᥈✲ࡋ ࡚࠸ࡃணᐃ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Okada, S. et al., Methods in Enzymology, 560, 331-353(2015) Suzuki, T. et al., Nature Protocols, 10, 715-732 (2015) Sakurai, M. et al., Genome Research, 24, 522-5 34(2014) - 392 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࣄࢺၚᾮ⣽⳦ྀෆᅾࡋࠊ⭠ෆ⅖㛵ࢃࡿ⣽⳦ࡢྠᐃࡑࡢᶵ⬟ࡢゎ᫂ ⱝᡭ◊✲㸦㹀㸧 㡲⏣ ற࣭᠕⩏ሿᏛ་Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࣄࢺࡢၚᾮ⣽⳦ྀࡣၚᾮඹᖖ⭠ෆ฿㐩ࡋࠊ ⭠ෆ⣽⳦࡞ࡾᚓࡿ⳦✀ࢆྵࡴࡇࡀᐃࡉࢀࡿࠋ ࣄࢺ⭠ෆ⣽⳦ྀ࠾࠸࡚ࡣࠊࡑࡢᵓᡂⓗ◚⥢ࡀ⅖ ᛶ⭠ᝈ(IBD)㛵㐃ࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡓࡀࠊ⏦ ㄳ⪅ࡽࡣࡇࢀࡲ࡛ࡢ◊✲࡛ IBD ᝈ⪅ࡢၚᾮ⣽⳦ྀࡀ ᖖ⪅ࡣࡁࡃ␗࡞ࡿᵓ㐀ࢆ♧ࡍࡇࢆⓎぢࡋࡓ (DNA Res., 21:15-25, 2014.)ࠋᮏ◊✲࡛ࡣࠊ௨ୖࡢ⫼ ᬒࢆ㋃ࡲ࠼ࠊࣄࢺၚᾮ⣽⳦ྀෆᅾࡋࠊ⭠ෆ⅖ 㛵ࢃࡿ⣽⳦ࡢ᥈⣴࣭ྠᐃࡑࡢᶵ⬟ࡢゎ᫂ࢆ┠ⓗ ࡍࡿࠋලయⓗࡣࠊIBD ᝈ⪅࠾ࡼࡧᖖ⪅ࡢၚᾮ⣽ ⳦ྀࢆ↓⳦࣐࢘ࢫᢞࡋࡓࣀࢺࣂ࣮࢜ࢺ࣐࢘ࢫ ࡛࠶ࡿࠕࣄࢺၚᾮ࣐࢘ࢫࠖࢆసᡂࡋࠊࡑࡢ⭠ෆ⣽⳦ ྀࡢゎᯒࡽࠊ ၚᾮ⣽⳦ྀෆᅾࡋ⭠ෆᐃ╔ྍ⬟ ࡞⣽⳦⩌ࢆ≉ᐃࡍࡿࠋࡲࡓࠊྛࣄࢺၚᾮ࣐࢘ࢫࡢ ⭠⢓⭷ᅛ᭷ᒙ࠾ࡅࡿ⅖ᛶ T ⣽⬊ࡢ㞟✚㔞ࢆゎᯒ ࡍࡿࡇ࡛ࠊ⅖㛵ࡍࡿ⣽⳦✀(⩌)ࢆྠᐃࡍࡿࠋ ࡉࡽࠊྠᐃࡉࢀࡓ⅖㛵㐃⣽⳦ࡢࢤࣀ࣒ゎᯒࢆ⾜ ࠺ࡇࡼࡗ࡚ࠊࡑࡢᶵ⬟ⓗ≉ᚩࢆ᫂ࡽࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࢤࣀ࣒ᨭ࡛ࡣࠊࣄࢺၚᾮ⣽⳦ྀࢆᢞࡋࡓ࣐࢘ࢫ ࡢ ⣅ ౽ ⣽ ⳦ ྀ ࡽ ᢳ ฟ ࡋ ࡓ ⳦ ྀ DNA ࡢ 16S rRNA 㑇ఏᏊ V1-2 㡿ᇦᑐࡋࠊMiseq ࡼࡿࣥ ࣉࣜࢥࣥࢩ࣮ࢡ࢚ࣥࢩࣥࢢ(࢚࣌ࣥࢻ)ࡢᨭࢆཷ ࡅࡿணᐃ࡛࠶ࡿࠋࡇࢀࡼࡾ 10,000~20,000 ࣮ࣜࢻ /᳨యࡢ 16S 㓄ิࢹ࣮ࢱࢆྲྀᚓࡋࠊ࣐࢘ࢫ⭠ෆᐃ╔ ࡋࡓ⳦✀ࡢ⤌ᡂࢆ᫂ࡽࡍࡿࡇࢆ┠ⓗࡍࡿࠋ ⣙ 150 ᳨యࡢ࣐࢘ࢫ⣅౽⣽⳦ྀࢆゎᯒࡍࡿࡇ࡛ࠊ ⅖㛵ࡍࡿ⣽⳦ࢆྠᐃࡋࠊ༢㞳ࢆヨࡳࡿࠋ ୖグࡢᕤ⛬ࢆ⤒࡚ࠊ༢ศ㞳ࡋࡓ⳦✀ࡽࢤࣀ࣒ DNA ࢆㄪᩚࡋࠊ ࢤࣀ࣒ᨭ࠾࠸࡚࣮࣍ࣝࢤࣀ࣒ࢩࣙࢵ ࢺ࢞ࣥࢩ࣮ࢡ࢚ࣥࢩࣥࢢࢆ⾜࠺ணᐃ࡛࠶ࡿࠋᮏィ⏬ ࡛ࡣࠊMiseq(࢚࣌ࣥࢻ)ࢆ⏝࠸ࠊ100~200 Mbase(ࢤ ࣀ࣒ࡢ x30~50 ⛬ᗘ)ࡢࢹ࣮ࢱࢆᚓࡿࡇࢆᐃࡋ࡚ ࠸ࡿࠋᚓࡽࢀࡓࢻࣛࣇࢺࢤࣀ࣒ࢹ࣮ࢱࡽࠊ⳦✀ࡢ ⣔⤫ࡸᶵ⬟ࡘ࠸࡚ゎ᫂ࡍࡿணᐃ࡛࠶ࡿࠋ ࠸࡞࠸≧ἣ࡛࠶ࡿࡀࠊᨭࡼࡗ࡚ୗグࡢᡂᯝࡀᚓ ࡽࢀࡿࡇࡀᮇᚅࡉࢀࡿࠋ ձ ࣄࢺၚᾮ⭠ෆ⣽⳦ࡢ┦㛵㛵ಀࡢゎ᫂ ղ ࣄࢺ⭠ෆ࠾ࡅࡿ⅖㛵ࡍࡿ⣽⳦ࡢ✀㢮࠾ ࡼࡧࡑࡢᛶ㉁⅖ㄏⓎ㛵ࡍࡿస⏝ᶵᗎࡢゎ ᫂ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡗ࡚ᖖᅾ⣽⳦ྀࡼࡿ⭠ෆ⅖ࡢస⏝ ᶵᗎࡀ᫂ࡽࡉࢀࡿࡇ࡛ࠊࡇࢀࡲ࡛ᑐⓗ࡞⒪ ἲࡋ↓ࡗࡓ⅖ᛶ⭠ᝈᑐࡍࡿࠊ✚ᴟⓗ࡞ ⒪ ἲࡢ㛤Ⓨ⧅ࡀࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ◊✲㐍⾜୰ࡢࡓࡵࠊᨭࡼࡿ⤖ᯝࡀࡲࡔᚓࡽࢀ࡚ - 393 - බເᨭㄢ㢟㸸2013 ᖺᗘ ⎔ቃ࣓ࢱࢤࣀ࣑ࢡࢫ࡛ᐩᰤ㣴ῐỈ†࡛ࡢ᪂ࡋ࠸❅⣲ᚠ⎔ീࢆ᥈ࡿ ᇶ┙◊✲㸦㸿㸧 ㄶゼ ⿱୍࣭୰ኸᏛ⌮ᕤᏛ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ᐩᰤ㣴†㸦ᾆ㸧࡛ anammox άᛶࡀ㧗࠸ᆅⅬ㸪 ᐇ㦂ᐊⓗ anammox ࡢάᛶࡀ⥔ᣢࡉࢀ࡚࠸ࡿࣂ࢜ ࣜࢡࢱ࣮ࡢᚤ⏕≀⩌ࡽᚓࡓ࣓ࢱࢤࣀ࣒ DNA ࡢ 㓄ิỴᐃࢆ⾜࠸㸪ࡑࢀࡒࢀࡢᚤ⏕≀⩌㞟ᵓ㐀❅⣲ ᚠ⎔ࢆᢸ࠺࡞ᶵ⬟㛵ࡍࡿ㑇ఏᏊ⩌ࡢᏑᅾ㔞 ⏤᮶⏕≀✀ሗࢆࡶ㸪ẚ㍑ゎᯒࡍࡿࠋ ࣓ࢱࢤࣀ࣒㓄ิ୰ࡢ㑇ఏᶵ⬟ண ࡣࠊࣜࣇࣞ ࣥࢫ࡞ࡿᚤ⏕≀ࢤࣀ࣒ࢹ࣮ࢱࡀᚲせ࡞ࡿࡀ㸪ᮏ ◊✲ࡢせ࡞ᑐ㇟࡛࠶ࡿ㸪anammox ᚤ⏕≀ࡢࢤࣀ࣒ ࢹ࣮ࢱࡣࡁࢃࡵ࡚ᑡᩘ࡛㸪ᑦୟࡘᡂࢤࣀ࣒ሗࡣ ⓙ↓࡛࠶ࡿࠋࡇࡢ≧ἣࢆᡴ㛤ࡍࡿࡓࡵ㸪ᾆ࡛ࡢඃ ༨ anammox ᚤ⏕≀࡛࠶ࡿ㸪Candidatus Brocadia ᒓࢆ ᐇ㦂ᐊ࡚㞟✚ࡋ㸪ࡇࡢ࣓ࢱࢤࣀ࣒ሗࡽ Brocadia ࢤࣀ࣒ࢆᵓ⠏ࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 1) Anammox 㞟 ✚ ᇵ 㣴 ࠾ ࡼ ࡧ ᾆ ᗏ Ἶ ࡢ total community DNA ࡢ࣓ࢱࢤࣀ࣒ゎᯒ㸸 㞟✚ᇵ㣴࣓ࢱ ࢤࣀ࣒⏝ total community DNA㸪1 ヨᩱ㸪࠾ࡼࡧᾆ ᗏἾ࣓ࢱࢤࣀ࣒ゎᯒ⏝ DNA ࢧࣥࣉࣝ 4 ヨᩱ㸦3 ᆅⅬ ࡼࡾ᥇ྲྀ㸧ࡘ࠸࡚㸪 MiSep ࡛ 1,300 ~ 3,500 paired end㸦ࣥࢧ࣮ࢺࢧࢬ 300~400 bp㸧࡛ሷᇶ㓄 ิࢆỴᐃࡋࡓࠋ࣓ࢱࢤࣀ࣒ࢹ࣮ࢱࡘ࠸࡚㸪࣮ࣜࢻ ࡢ R1, R2 ࢭࣥࣈࣝࢆ⾜ࡗ࡚ᚓࡽࢀࡓ 400bp ௨ୖࡢ 㓄ิࢆࠊศᢸ◊✲⪅ࡢ㧘ぢࡀ⾜࠺㑇ఏᏊ㡿ᇦ᥎ᐃ㸪 MAPLE (Metabolic And Physiological potentiaL Evaluator)ࢩ ࢫࢸ࣒ࡼࡿ⏕⌮࣭௦ㅰᶵ⬟ゎᯒ౪ヨࡋࡓࠋࡇࢀ ࡣูࠊ࣓ࢱࢤࣀ࣒୰ࡢ 16S rDNA ࢹ࣮ࢱᇶ࡙ࡃ 㛛࠾ࡼࡧ⛉࡛ࣞ࣋ࣝࡢ⏕≀✀ศ㢮ࢆ⾜ࡗࡓࠋ 2) 㞟 ✚ ᇵ 㣴 ⣔ ࠾ ࡅ ࡿ ඃ ༨ ✀ ࡢ ࢤ ࣀ ࣒ ᵓ ⠏ 㸸 Anammox ᚤ⏕≀ࡢ㞟✚ᇵ㣴ࢆ☜❧ࡋ㸪ࡑࡢ total community DNA ࢆᢳฟࡋࡓࠋ࣓ࢱࢤࣀ࣒ DNA ࡽ PacBio ࢆ⏝࠸࡚ anammox ᚤ⏕≀ࢆࡣࡌࡵࡍࡿඃ༨ ✀ࡢࢤࣀ࣒ᵓ⠏ࢆᐇࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 1) Anammox 㞟 ✚ ᇵ 㣴 ࠾ ࡼ ࡧ ᾆ ᗏ Ἶ ࡢ total community DNA ࡢ࣓ࢱࢤࣀ࣒ゎᯒ㸸 ࣭16S rDNA ࢆࡶ㸪ྛヨᩱ࠾ࡅࡿ⏕≀✀ศ㢮ࢆ ⾜ࡗࡓࠋࡑࡢ⤖ᯝ㸪anammox 㞟✚ᇵ㣴㸪࠾ࡼࡧ ᾆሁ✚≀࠸ࡎࢀ࠾࠸࡚ࡶඹ㏻ࡋ࡚㸪Firmicutes, Proteobacteria, Chloroflexi ࡢ 3 㛛ᒓࡍࡿ⏕≀✀ࡀ 㢖ᗘ㧗ࡃ᳨ฟࡉࢀࡓࠋAnammox 㞟✚ᇵ㣴࡛ࡣ㸪 anammox ⳦ࡀᒓࡍࡿ Planctomycetes 㛛ࡢ⏕≀✀ࡀண ㏻ࡾ㧗࠸㢖ᗘ᳨࡛ฟࡉࢀࡓࠋ ୍࣭᪉ࠊ16S rDNA ࡣࠊ⏕≀✀ࡼࡾࡑࡢࢥࣆ࣮ᩘࡀ ࡲࡕࡲࡕ࡞ࡓࡵࠊ⏕≀✀⤌ᡂࡢ᥎ ⏝࠸ࡿࡇࡀ ࡛ࡁ࡞࠸ࠋࡑࡇ࡛ࠊMAPLE ࢩࢫࢸ࣒ࡼࡾ᳨ฟࡉ ࢀࡓࣜ࣎ࢯ࣮࣒ࢱࣥࣃࢡ㉁㑇ఏᏊࡢ⏤᮶⏕≀✀⤌ ᡂࡽࠊྛ࣓ࢱࢤࣀ࣒ࢧࣥࣉࣝࡢ㛛࡛ࣞ࣋ࣝࡢ⏕≀ ✀⤌ᡂࢆࡾฟࡋࡓࠋࣜ࣎ࢯ࣮࣒ࢱࣥࣃࢡ㉁ࡢ⏤᮶ ⏕≀✀ࡽ anammox ᚤ⏕≀ࡢయ༨ࡵࡿྜࢆ ㄪࡓࡇࢁࠊ᭱ࡶ anammox άᛶࡢ㧗࠸ᆅⅬ࡛ࡣࠊ యࡢ 0.3㸣ࠊ᭱ࡶప࠸ᆅⅬ࡛ࡣ 0.08%ぢ✚ࡶࡽ ࢀࡓࠋࡇࡢ⤖ᯝࡽࠊanammox άᛶࡢ⛬ᗘࡣࠊ anammox ᚤ⏕≀ࡢᏑᅾ㔞౫Ꮡࡍࡿ⪃࠼ࡽࢀࡓࠋ 2) 㞟✚ᇵ㣴ࡉࢀࡓ anammox ᚤ⏕≀ࡢࢤࣀ࣒ᵓ⠏㸸 PacBio ࢆ⏝࠸࡚㸪anammox ᚤ⏕≀࡛࠶ࡿ㸪Candidatus Brocadia ᒓࡢࢤࣀ࣒ᵓ⠏ࢆ⾜ࡗࡓࠋ⌧ᅾ᭱⤊☜ㄆ ࢆ⾜ࡗ࡚࠸ࡿࡀ㸪ࡇࢀࡲ࡛࡛᭱ᑠࡢ contig ᩘ 2 ࡲ࡛ ࢤࣀ࣒ࡀᵓ⠏ࡉࢀࡓࠋ୍᪉ࠊࡇࢀྠඃ༨✀ ࡢ୍ࡘ࡛࠶ࡿ Armatimonadetes 㛛ࡢࢤࣀ࣒ࡀᵓ⠏ࡉ ࢀࡓࡢ࡛ࠊ⌧ᅾ࣐ࢽࣗࣝࣀࢸ࣮ࢩࣙࣥࢆᐇࡋ ࡚࠸ࡿࠋ 3) ࣓ࢱࢤࣀ࣒ࡢᶵ⬟࣏ࢸࣥࢩࣕࣝゎᯒ㸸 Anammox ᚤ⏕≀ඹᏑ⳦ࡢ㛵ಀࢆ᥎ ࡍࡿᡭࡀࡾࢆᚓࡿ ࡓࡵ㸪ྲྀᚓࡉࢀࡓ࣓ࢱࢤࣀ࣒ሗࡽ MAPLE ࢩࢫ ࢸ࣒ࢆ⏝࠸࡚௦ㅰᶵ⬟࣏ࢸࣥࢩࣕࣝࢆྍどࡋࡓࠋ ࡇࡢゎᯒࡼࡗ࡚㸪ḟࡢ⤖ᯝࡀᚓࡽࢀࡓࠋ Anammox άᛶࡀ᭱ࡶ㧗࠸ᆅⅬ࡛ࡣ㸪㏫◪⬺ ❅ࡢάᛶࡀపࡗࡓࠋྠᆅⅬ࡛ࡣ㸪◪⬺❅ࡢ௦ ㅰᶵ⬟ࣔࢪ࣮ࣗࣝࡢ abundance ࡶపࡗࡓࠋ୍᪉㸪 anammox άᛶࡀ᭱ࡶప࠸ᆅⅬ࡛ࡣࠊ◪⬺❅ࡢά ᛶ࠾ࡼࡧ௦ㅰᶵ⬟ࣔࢪ࣮ࣗࣝ abundance ࡢ࠸ࡎࢀࡶ ࡀ㧗ࡗࡓࠋanammox άᛶࡀୖグ㸰ᆅⅬࡢ୰㛫ࡢᆅ Ⅼ࡛ࡣ Betaproteobacteria ࠾ࡼࡧ Gammaproteobacteria ࡀ⬺❅ᛂࢆᢸ࠸㸪⯆῝࠸ࡇ㸪ୖグ 2 ᆅⅬ࡛ ࡣࡇࢀࡲ࡛⬺❅⳦ࡢศ㞳ࡢ࡞࠸ Deltaproteobacteria ࡀ⬺❅⬟ࢆᣢࡘྍ⬟ᛶࡶ♧၀ࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 1) Anammox 㞟 ✚ ᇵ 㣴 ࠾ ࡼ ࡧ ᾆ ᗏ Ἶ ࡢ total community DNA ࡢ࣓ࢱࢤࣀ࣒ゎᯒ㸸 ࣭࣓ࢱࢤࣀ࣒ሗࡽ anammox ᚤ⏕≀ࡀ⎔ቃ୰༨ ࡵࡿྜࢆ᥎ ࡋ㸪anammox άᛶᕪࡀぢࡽࢀࡿ ⎔ቃ୰࠾ࡅࡿᚤ⏕≀⩌㞟ᵓ㐀ࡢᐇീ㏕ࡿࡇ ࡀ࡛ࡁࡓࠋ ࣭ῐỈ† anammox ᚤ⏕≀ࡢ㞟✚ᇵ㣴⣔࡛㸪ᚤ ⏕≀⩌㞟ᵓ㐀ඹ㏻ᛶぢࡽࢀ㸪anammox ᚤ⏕≀ ࡑࡢࡢᚤ⏕≀ࡢ㛵ಀࡣ㸪࠶ࡿᬑ㐢ᛶࡀᏑᅾࡍ ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ 2) 㞟✚ᇵ㣴ࡉࢀࡓ anammox ᚤ⏕≀ࡢࢤࣀ࣒ᵓ⠏㸸 ᮏ◊✲࠾࠸࡚ࠊanammox ᛂࢆᢸ࠺ Brocadia ඹ Ꮡ⳦ࡋ࡚▱ࡽࢀࡿ Armarimonadetes 㛛ࡢᮍᇵ㣴⳦ ࡢࢤࣀ࣒ሗࡀᚓࡽࢀࡓࡇ࡛ࠊ㞟✚ᇵ㣴⣔࠾ࡅ ࡿ⏕ែ⣔ᵓ⠏ཎ⌮ࡢゎ᫂ࡸ⎔ቃ୰࠾ࡅࡿ anammox ⳦ࡢᙺࡢゎ᫂ྥࡅࠊࡁ࡞୍Ṍࢆ㋃ࡳฟࡋࡓ ⪃࠼ࡽࢀࡿࠋᚋࡇࢀࡽࡢࢤࣀ࣒ሗ࣓ࢱࢤࣀ࣒ ሗࢆ⥲ྜⓗゎᯒࡍࡿࡇ࡛ࠊ❅⣲௦ㅰ࠾ࡅࡿ anammox ࡢព⩏ࡸ anammox ᚤ⏕≀ࡢᐇࡏࡲࢀࡿ ࡶࡢᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ㸦⌧ᅾ‽ഛ୰㸧ࠋ - 394 - බເᨭㄢ㢟㸸2012㹼2013 ᖺᗘ ༡ᴟịᗋᗏ㠃ࡽ᳨ฟࡉࢀࡓ 72 ᖺ௨ୖ๓ࡢᚤ⏕≀᥈ᰝ ⱝᡭ◊✲㸦㹀㸧 ℩ᕝ 㧗ᘯ࣭ሗ࣭ࢩࢫࢸ࣒◊✲ᶵᵓ ᪂㡿ᇦ⼥ྜ◊✲ࢭࣥࢱ࣮㸪ᅜ❧ᴟᆅ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ༡ᴟịᗋࢻ࣮࣒ࡩࡌᇶᆅ࡛ࡢࢫࢥ᥀๐ࡼ ࡾ㸪༡ᴟịᗋୗࡢịヨᩱᒾ┙⏤᮶ࡢ⢏≧≀ヨᩱࡀ ᚓࡽࢀࡓ㸬ࡇࢀࡽࡣᑡ࡞ࡃࡶ 72 ᖺ௨ୖ๓ᑒࡌ ㎸ࡵࡽࢀࡓྍ⬟ᛶࡀᙉࡃ㸪༡ᴟịᗋୗࡢ⎔ቃࢆ᥎ᐃ ࡉࢀ࠺ࡿヨᩱࡢ୰࡛㸪᭱ࡶඃࢀࡓヨᩱࡢ୍ࡘ࡛࠶ࡿ ゝ࠼ࡿ㸬ᮏ◊✲ࡢ┠ⓗࡣ㸪ࡇࡢ≉␗࡞ᴟ㝈⎔ቃ ࡽᚓࡽࢀࡓᚤ⏕≀ࢆゎᯒࡋ㸪ࡑࡢᶵ⬟ࡢゎ᫂ࡽ㸪 ࠕே㢮ṧࡉࢀࡓ᭱ᚋࡢࣇࣟࣥࢸࡢࡦࡘࠖ ࡉࢀࡓ༡ᴟịᗋୗࡢ⏕ែ⣔ࡸ⎔ቃࢆゎࡁ᫂ࡍࡇ ࡛࠶ࡿ㸬ࡇࢀࡼࡾᚤ⏕≀ࡢ⏕⌮࣭⏕ែࢆゎ᫂ࡍࡿࡇ ࡣ㸪ே㢮ࡗ࡚ᮍ▱࡛࠶ࡿᴟ㝈⎔ቃୗ࠾ࡅࡿ ⏕ែ⣔ࡢᐇయᡂ❧㐣⛬ࡢゎ᫂㸪㔜せ࡞ᙺࢆᯝ ࡓࡍ⪃࠼ࡽࢀࡿ㸬ࡋࡋ㸪⌧Ꮡࡋ࡚࠸ࡓࡋ࡚ࡶ ྠ⣽⳦ࡢศ㞳ᇵ㣴ࡣప ࣭㧗ᅽୗࡢᴟ㝈⎔ቃ⏕ᜥ ࡋ࡚࠸ࡿࡇ㸪ヨᩱᖺ௦ࡀྂ࠸ࡇࡽᅔ㞴ࡀண ࡉࢀ㸪ࡢࡇࢁᡂຌࡋ࡚࠸࡞࠸㸬 ᮏ◊✲࡛ࡣ㸪ᇵ㣴⬟⣽⳦ࡢᑡᩘ⣽⬊ࡽࡢࢤࣀ ࣒㛗ྲྀᚓἲࢆᛂ⏝ࡋ࡚㸪ᇵ㣴ࢆࡉࡎࢤࣀ࣒ ゎᯒࢆ⾜࡞࠸㸪ࡑࡢᶵ⬟ゎ᫂ࡽࡇࢀࡲ࡛ᮍ▱࡛࠶ ࡗࡓ༡ᴟịᗋୗࡢ⏕ែ⣔ࡢ᥎ᐃࢆヨࡳࡓ㸬 㸺ᨭࡢෆᐜ㸼 ༡ᴟࡢịᗋᗏ㠃ࡽ᥇ྲྀࡉࢀࡓヨᩱ࠾ࡼࡧ㸪༡ᴟ ịᗋᗏ㒊ࡢ⎔ቃẚ㍑ࡍࡿࡓࡵࢫࢥヨᩱ ࡽࡢ㸪ࢤࣀ࣒ࣛࣈ࣮ࣛࣜࡢసᡂ࠾ࡼࡧࢤࣀ࣒㓄ิ ゎㄞࡢᨭࢆ࠾ࡇ࡞ࡗ࡚㡬࠸ࡓ㸬ịἙヨᩱ୰ࡢ⣽⬊ ᐦᗘࡣప࠸ࡓࡵ㸪DNA ᢳฟ⏘≀ࡸᚤ㔞⣽⬊ࡽ㸪 phi29 ࢆ⏝࠸ࡓᑡᩘ⣽⬊ࡽࡢࢤࣀ࣒ቑᖜᡭἲࡸ㸪 ྂ௦ヨᩱࡽࡢ DNA ࣛࣈ࣮ࣛࣜᵓ⠏ඃࢀࡓ Ancient Illumina sequencing library ἲࢆ⏝࠸࡚ࢤ ࣀ࣒ቑᖜࢆ࠾ࡇ࡞ࡗࡓ㸬ቑᖜࣂࢫ࡞ࡢᙳ㡪ࢆ పῶࡍࡿࡓࡵ㸪ྠ୍ DNA ࢧࣥࣉࣝࢆศࡅ࡚㸪㸳ᮏ⊂ ❧ࢤࣀ࣒ቑᖜࢆ࠾ࡇ࡞࠸ゎᯒ⏝ࢧࣥࣉࣝࡋࡓ㸬 ࡇࢀࡽࡢࢧࣥࣉࣝ㛵ࡋ࡚㸪ࢤࣀ࣒ᨭάືࡼࡾ㸪 ࢤࣀ࣒ࣛࣈ࣮ࣛࣜసᡂࡸ࣑ࣝࢼࡢ Hiseq ࠾ࡼࡧ Miseq ࢩ࣮ࢡ࢚ࣥࢧ࣮ࢆ⏝࠸ࡓ㑇ఏᏊゎㄞࢆ࠾ࡇ࡞ ࡗ࡚㡬࠸ࡓ㸬 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ༡ᴟịᗋᗏ㒊ヨᩱࡢ࣓ࢱࢤࣀ࣒ゎᯒࡼࡾᚓࡽࢀ ࡓ㓄ิᑐࡋ࡚┦ྠᛶ᳨⣴ࢆ࠾ࡇ࡞࠸㸪ࢱ࣮ࢤࢵࢺ ࡢ㑇ఏᏊ㓄ิ⏤᮶⪃࠼ࡽࢀࡿ࣮ࣜࢻᢳฟࡸᚤ⏕≀ ⩌ࡢ⣔⤫᥎ᐃ㸪㑇ఏᏊ⩌ࡢᶵ⬟ࡢゎᯒࢆ࠾ࡇ࡞ࡗࡓ㸬 ࡑࡢ⤖ᯝ㸪㸰㸮㸮ࡽ㸲㸮㸮✀㢮ࡢከᵝ࡞ᚤ⏕≀⩌ ࢆ᳨ฟࡍࡿࡇࡀ࡛ࡁࡓ㸬⯆῝࠸ࡇịᗋୗ ࡽᚓࡽࢀࡓ࣓ࢱࢤࣀ࣒㓄ิࢆゎᯒࡋࡓ⤖ᯝ㸪ࢩࣀ ࣂࢡࢸࣜࢆྵࡴྛ✀ࣂࢡࢸࣜ࠾ࡼࡧ࣮࢟࡞ ࡀ᳨ฟࡉࢀ,ࢻ࣮࣒ࡩࡌᇶᆅࡽᚓࡽࢀࡓ༡ᴟịᗋ ୗヨᩱࡣከᵝ࡞ᚤ⏕≀⩌ࡀᏑᅾࡋ࡚࠸ࡿࡀ᫂ࡽ ࡞ࡗࡓ㸬ࡇࢀࡽࡢᚤ⏕≀ࡣ㸪ࡇࢀࡲ࡛ࡢศᯒ⤖ ᯝࡣ㧗࠸┦ྠᛶࢆ♧ࡉࡎ㸪⌧ᅾㄪࡽࢀ࡚࠸ࡿ༡ ᴟࡢ㞷ịࡸᅵተ㸪†➼ࡢ⎔ቃࡣ⏕ᜥࡋ࡚࠸࡞࠸ ࡇࡀศࡗࡓ㸬ྠ⣽⳦⩌ࡣ᪤▱ࡢࡶࡢࡣࡁࡃ ␗࡞ࡿ㓄ิࢆᣢࡘࡇࡽ㸪ࡇࢀࡣ༡ᴟịᗋୗ࡞ ࡢᴟ㝈⎔ቃ㐺ᛂࡋ࡚࠸ࡿ≉␗࡞⣔⤫࡛࠶ࡿࡇࡀ ♧၀ࡉࢀࡓ㸬༡ᴟịᗋୗࡢ⎔ቃ᥎ᐃࢆ࠾ࡇ࡞࠺ࡓࡵ ࢤࣀ࣒ሗࡣ㠀ᖖ᭷ຠ࡛࠶ࡿࡇࡀศࡾ㸪ࡇ ࢀࡲ࡛᥎ᐃᅔ㞴࡛࠶ࡗࡓịᗋୗ⎔ቃࡢ≀⌮Ꮫⓗ࡞ ᛶ㉁ண ࡢ㈉⊩ࡶ⧅ࡀࡿணࡉࢀࡿ㸬 ࡋࡋ࡞ࡀࡽ㸪ศᯒࢆᐇᆅࡋࡓᅜ❧ᴟᆅ◊✲ᡤࡢ ࢡ࣮࣮࣒ࣜࣥࣝࡀᨾ㞀ࡋ࡚࠸ࡿࡇࡀุ᫂ࡋ㸪ịᗋ ୗࡢᚤ⏕≀࠸࠺⣽⬊ᩘࡀᕼⷧࡘヨᩱᖺ௦ࡀྂ࠸ ヨᩱᑐࡋ࡚㸪༑ศ࡞ࢡ࣮ࣜࣥ࡞⎔ቃୗ࡛ศᯒࢆ㐙 ⾜ࡍࡿࡇࡀ࡛ࡁ࡞ࡗࡓ㸬ࢫࢥࡣᖺ௦ࡢྂ ࠸ヨᩱ࡛࠶ࡿࡓࡵ DNA ࡢ≧ែࡀ㠀ᖖᝏࡃ㸪ࡑࡢࡓ ࡵࢤࣀ࣒ࣛࣈ࣮ࣛࣜ୰ࡣࣂࢵࢡࢢࣛ࢘ࣥࢻ࡛࠶ ࡿᐇ㦂ᐊ࡞ࡽࡢࢥࣥࢱ࣑ࢿ࣮ࢩࣙࣥ㓄ิࡀከࡃ ΰᅾࡋ࡚࠸ࡿ㸬⌧ᅾ㸪ࡇࢀࡽࡢᚤ⏕≀✀ࡢࢤࣀ࣒ ࢥ࣮ࢻࡉࢀ࡚࠸ࡿ㑇ఏᏊ⩌ࡢᶵ⬟ศᕸࡣ㸪ሗ࣭ࢩ ࢫࢸ࣒◊✲ᶵᵓ ᪂㡿ᇦ⼥ྜ◊✲ࢭࣥࢱ࣮ࡢ㏆⸨ఙ ≉௵◊✲ဨࡀ㐍ࡵ࡚࠸ࡿẁ㝵࡛࠶ࡿ㸬 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ༡ᴟịᗋᗏ㒊ヨᩱࡽࡢ࣓ࢱࢤࣀ࣒ゎᯒຍ࠼㸪 ᚤ㔞⣽⬊ࡽࡢࢤࣀ࣒ࢩ࣮ࢣࣥࢩࣥࢢࢆ࠾ࡇ࡞࠸㸪 ༡ᴟịᗋୗ࠸࠺ᴟ㝈⎔ቃ࠾ࡅࡿ⏕ែ⣔ࡢᐇయ ㏕ࡿࡇࡀ࡛ࡁࡓ㸬ࡲࡓ㸪16S rRNA 㑇ఏᏊࡢࡳ࡞ ࡽࡎ㸪」ᩘࡢᶵ⬟㑇ఏᏊ⩌ࡢ᳨ฟᡂຌ࡛ࡁࡓࡇ ࡣ㸪ịᑒࡌ㎸ࡵࡽࢀࡓኴྂࡢᚤ⏕≀ࡢゎ᫂ࢆྍ⬟ ࡍࡿࡇࡀᮇᚅࡉࢀࡿ㸬 ᮏ◊✲ࡣ㸪ྂ௦ヨᩱࡽࡢᚤ㔞 DNA ࡢቑᖜࡸࢤࣀ ࣒ゎㄞࢆ࠾ࡇ࡞࠺ࡇ࡛㸪ヨᩱᖺ௦ࡢྂ࠸ヨᩱࡽ ࡢࢤࣀ࣒ゎᯒࢆ࠾ࡇ࡞࠺ࡇᡂຌࡋࡓ㸬ᚋ㸪ị ヨᩱࡢࡳ࡞ࡽࡎ༤≀㤋࡞㈓ⶶࡉࢀ࡚࠸ࡿᵝࠎ࡞ ྂ௦ヨᩱࡽࡢࠕྂ௦ DNA ◊✲ࠖᛂ⏝࡛ࡁࡿࡇ ࡀᮇᚅࡉࢀࡿ㸬 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 395 - බເᨭㄢ㢟㸸2010 ᖺᗘࠊ2012 ᖺᗘࠊ2015 ᖺᗘ ࣄ࣓࣑࢝ࢷ࢟ࣔࡢᛶࣇ࢙ࣟࣔࣥཷᐜయྠᐃ≉ᛶゎᯒ ᇶ┙◊✲㸦㹁㸧 ࣄ࣓࣑࢝ࢷ࢟ࣔࡢᛶᰁⰍయ㡿ᇦゎᯒࡼࡿ⏕Ṫᵝᘧ㐍ࡢゎ᫂ ᇶ┙◊✲㸦㹀㸧 㛵ᮏ ᘯஅ࣭᪥ᮏዪᏊᏛ⌮Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ᮏ◊✲࡛ὀ┠ࡋࡓ༢⣽⬊᥋ྜ⸴㢮࡛࠶ࡿࣄ࣓࣑࢝ ࢷ࢟ࣔࡣࠊ㝣ୖ᳜≀ࡶࡗࡶ㏆⦕࡞༢⣽⬊⸴㢮࡛ ࠶ࡿࡇࡀᨭᣢࡉࢀ࡚࠾ࡾࠊࡲࡓ㑇ఏᏊᑟධ⣔ࡶ☜ ❧ࡋࠊ᳜≀ࡢ᭷ᛶ⏕Ṫࡢᇶᮏ㒊ศゎ᫂ࡢࡓࡵࡢࣔࢹ ࣝ⏕≀⨨࡙ࡅࡽࢀࡿࠋࡇࢀࡲ࡛ࠊ㑇ఏⓗ␗࡞ ࡿ㸩ࠊ㸫ᆺ⣽⬊ࡢ㛫࡛᭷ᛶ⏕Ṫࡋ࡚ࠊ᥋ྜᏊࢆᙧᡂ ࡍࡿ࣊ࢸࣟࢱࣜࢵࢡᰴ㸦Ṫ⣔⤫㸧ࢆ⏝࠸ࡓ◊✲ࢆ 㐍ࡵ࡚࠾ࡾࠊ᭷ᛶ⏕ṪࡲࡓࡣᛶⓎ⌧㛵㐃ࡍࡿ ✀ ࡢ㑇ఏᏊ⩌ࡀ᫂ࡽࡉࢀ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽࠊ ࡇࢀࡽࡣ࠸ࡎࢀࡶ୧᥋ྜᆺࢤࣀ࣒Ꮡᅾࡋ࡚࠸ࡿ㑇 ఏᏊ࡛࠶ࡾࠊࡇࢀࡽࢆ⤫ᚚࡋ࡚࠸ࡿ᥋ྜᆺ≉␗ⓗ࡞ ࢤࣀ࣒㡿ᇦࠊᛶỴᐃ㑇ఏᏊࡢᐇయࡘ࠸࡚ࡣ࡛᫂ ࠶ࡗࡓࠋ ࡑࡇ࡛ᮏ◊✲࡛ࡣࠊࣄ࣓࣑࢝ࢷ࢟ࣔ㸩ᆺѸᆺࡑࢀ ࡒࢀࡢ⣔⤫ಖᏑᰴࡘ࠸࡚ࠊ᪂ᆺࢩ࣮ࢡ࢚ࣥࢧ࣮ࢆ ⏝࠸࡚ࠊᴫせࢤࣀ࣒㓄ิࢆỴᐃࡍࡿࡇࡋࡓࠋࢤ ࣀ࣒ࢭࣥࣈࣝࡢᚋࠊࡇࢀࡲ࡛ᚓ࡚࠸ࡿ∦᪉ࡢ᥋ ྜᆺ≉␗ⓗᏑᅾࡍࡿ㑇ఏᏊࡢ࣐ࢵࣆࣥࢢࢆ㊊ࡀ ࡾࡋ࡚ࠊ୧ࢤࣀ࣒ࡢẚ㍑ࢆ⾜࠸ࠊᛶᰁⰍయ㡿ᇦࡢ ≉ᐃࠊᛶỴᐃ㑇ఏᏊࡢྠᐃࢆ┠ᣦࡍࡇࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 㸯㸬ࣄ࣓࣑࢝ࢷ࢟ࣔࡢ㸩ᆺ⣽⬊㸦1,(6 ᰴ㸧ࠊѸᆺ⣽ ⬊㸦1,(6 ᰴ㸧ࡑࢀࡒࢀࡢ⣔⤫ಖᏑᰴࡘ࠸࡚ࠊ +L6(4 ࢆ⏝࠸࡚ࠊNE ࡢ SDLUHGHQGOLEUDU\ࠊNENENE ࡢ PDWHSDLU OLEUDU\ ࢆỴᐃࡋࡓࠋ 㸰㸬ࣄ࣓࣑࢝ࢷ࢟ࣔࡢ㸩ᆺ⣽⬊㸦1,(6 ᰴ㸧ࠊѸᆺ⣽ ⬊㸦1,(6 ᰴ㸧ࡑࢀࡒࢀࡢ⣔⤫ಖᏑᰴࡘ࠸࡚ࠊ%$& ࣛࣈ࣮ࣛࣜࢆస〇ࡋࡓࠋ 㸱㸬ࣄ࣓࣑࢝ࢷ࢟ࣔࡢᰤ㣴ቑṪᮇࠊ᭷ᛶ⏕Ṫᮇࡢィ ✀ࡢࢫࢸ࣮ࢪࡽㄪ〇ࡋࡓ 51$ ࡘ࠸࡚ࠊ+L6(4 ࢆ⏝࠸ࡓ WUDQVFULSWRPH ゎᯒࢆ⾜ࡗࡓࠋ 㸲㸬1,(6 ᰴࠊ1,(6 ᰴࡑࢀࡒࢀࡘ࠸࡚ࠊ3& ࢣ࣑ࢫࢺ࣮ࣜࢆ⏝࠸ࡓ 3DF%LR ࡼࡿࢤࣀ࣒ࢩ࣮ࢡ ࢚ࣥࢫࢆ⾜ࡗࡓࠋ 㸳㸬3DF%LR ཬࡧ ,OOXPLQD ࢹ࣮ࢱࡢࣁࣈࣜࢵࢻࢭ ࣥࣈࣜࢆ⾜࠺ࡓࡵࡢࠊࣄ࣓࣑࢝ࢷ࢟ࣔ⏝ゎᯒࣝࢦ ࣜࢬ࣒タィࢆ㐍ࡵࡓࠋ 㸴㸬ࣄ࣓࣑࢝ࢷ࢟ࣔࡢ㓄ᏊᏞᰴ㸦 ᰴ㸧ࡢ࣑ࣝ ࢼ LQGH[ ࢩ࣮ࢣࣥࢫࢆ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 $+,6(4 ࡼࡿࢤࣀ࣒ゎᯒ ࣄ࣓࣑࢝ࢷ࢟ࣔࡢᴫせࢤࣀ࣒ゎᯒࡘ࠸࡚ࡣࠊ ᅜ❧㑇ఏᏛ◊✲ᡤ࣭㇏⏣ඛ⏕ࡢࡶࠊ+L6(4 ࢆ⏝ ࠸࡚ࠊ㸩ᆺ࠾ࡼࡧѸᆺࡑࢀࡒࢀࡢ㓄ิࡀỴᐃࡉࢀࡓࠋ NPHU 㢖ᗘゎᯒࡼࡿࢤࣀ࣒ࢧࢬ᥎ᐃ࡛ࡣ 0E ࡞ࡗࡓࠋ୍᪉ࠊ$//3$7+6/* ࡼࡿゎᯒ࡛ࠊ㸩 ᆺ࠾ࡼࡧѸᆺࢤࣀ࣒ࡢ &RQWLJ1 ࡀ NE ཬࡧ NE ࠊVFDIIROG1 ࡀ NE ཬࡧ NE ࡞ࡾࠊ VFDIIROG 㛗ࡢྜィࡣ⣙ 0E ࡞ࡗࡓࠋVFDIIROG ࡣ༑ศࡘ࡞ࡀࡗ࡚࠸࡞࠸⌮⏤ࡋ࡚ࠊࣄ࣓࣑࢝ࢷ ࢟ࣔࡢࢤࣀ࣒࠾࠸࡚つᶍ࡞ ࠥ ࢥࣆ࣮㡿ᇦࡀᏑ ᅾࡋ࡚␗࡞ࡿከᆺࡀ࠶ࡿ࠸࠺」㞧࡞⌧㇟ࡀ㛵㐃ࡋ ࡚࠸ࡿ⪃࠼ࡽࢀࡓࠋNPHU ࢢࣛࣇ࡛㸰ಸࡽ㸱ಸ ┦ᙜࡍࡿࣆ࣮ࢡࡀㄆ㆑࡛ࡁࠊࡉࡽࠊVFDIIROG ᆺࠊ̾ᆺࡑࢀࡒࢀࡢ࣮ࣜࢻࢹ࣮ࢱࢆ࣐ࢵࣆࣥࢢࡋࡓ ࡁࡢཌࡳࡢẚࡀࠊ ࡞ࡿ㡿ᇦࡀከᩘぢࡽࢀ ࡿࡇࡽࠊ⣔⤫ࡼࡗ࡚ࡑࡢࢥࣆ࣮ᩘࡶ␗࡞ࡗ࡚ ࠸ࡿ⪃࠼ࡽࢀࡿࠋ≧ἣࡣࠊ࣊ࢸࣟ᥋ྜᗘࡢ㧗࠸ ಸయࢤࣀ࣒ఝ࡚࠸ࡿࡢ࡛ SODWDQXV ࢆヨࡳࡓࡀࠊ ࢭࣥࣈ࣮ࣜࡀ⤊ࡋ࡞ࡗࡓࠋ %㸩ᆺ࠾ࡼࡧѸᆺ⣽⬊ࡽࡢ %$& ࣛࣈ࣮ࣛࣜస〇 ⌮Ꮫ◊✲ᡤ࣭㯮ᮌඛ⏕ࡼࡾࠊྛ⣔⤫ᰴࡢ %$& ࣛࣈ࣮ࣛࣜࡀస〇ࡉࢀࡓࠋѸᆺ⣽⬊ࡢ %$& ࣛࣈࣛ ࣮ࣜࡣᖹᆒ⣙ NE ࣥࢧ࣮ࢺࠊZHOOSODWH ࡛ ᯛศࠊ㸩ᆺ⣽⬊ࡢ %$& ࣛࣈ࣮ࣛࣜࡣᖹᆒ⣙ NE ࣥࢧ࣮ࢺࠊZHOOSODWH ࡛ ᯛศ࡞ ࡗࡓࠋ & 7UDQVFULSWRPH ゎᯒ ⏕ά⎔ࡢᵝࠎ࡞ࢫࢸ࣮ࢪࡽ ᮲௳ࢆ㑅ࡧ 51$ ࢆㄪ〇ࡋࠊᮾ᪂㡿ᇦ࣭㕥ᮌ✨ඛ⏕ࡢᨭࡼࡾ WUDQVFULSWRPH ࢹ࣮ࢱࢆྲྀᚓࡋࡓࠋ7ULQLW\ ࡼࡿ ࢭࣥࣈࣝࡢ⤖ᯝࠊFRQWLJ ࡀᚓࡽࢀࠊ⌧Ⅼ࡛ ࡣ࡞ࡾከࡃࡢ LVRIRUP ࢆྵࡴࡇࡀ♧၀ࡉࢀࡓࠋ ࡲࡓࡇࡢ⤖ᯝࡣࠊࢤࣀ࣒ゎㄞࡢ⤖ᯝࡽ᥎ᐃࡉࢀࡓ ࢤࣀ࣒ෆつᶍ࡞ ࠥ ࢥࣆ࣮㡿ᇦࡀᏑᅾࡋ࡚␗࡞ ࡿከᆺࡀ࠶ࡿ≧ἣࢆᫎࡋ࡚࠸ࡿࡶ⪃࠼ࡽࢀࡓࠋ ᚓࡽࢀࡓ FRQWLJ 㓄ิࡽࠊѸᆺ࡛≉␗ⓗⓎ⌧ࡍ ࡿ 㓄ิࢆ⤠ࡾ㎸ࡳࠊѸᆺࢤࣀ࣒ࡢࡳᏑᅾࡍࡿ ᛮࢃࢀࡿ 㓄ิࢆ⤠ࡾ㎸ࢇࡔࠋ㸩ᆺѸᆺࢆࡅྜࢃ ࡏ࡚ᏊᏞᰴࢆྲྀᚓࡋࠊ㐃㙐ゎᯒࢆ⾜ࡗࡓࡇࢁࠊ㸯 ࡘࡢ㓄ิࡢࡳࡀ㸫ᆺࡢᏊᏞᰴ㐃㙐ࡋ࡚࠸ࡓࠋ ࡇࡢ㓄ิࢆᣢࡘ㑇ఏᏊࡣ㌿ᅉᏊࢆࢥ࣮ࢻࡋ࡚࠾ࡾࠊ 㸫ᆺ⣽⬊≉␗ⓗᏑᅾࡋ࡚࠸ࡿࡇࡽࠊ&S0LQXV ྡࡅࡽࢀࡓࠋࡇࡢ㑇ఏᏊࢆᣢࡓ࡞࠸㸩ᆺ⣽⬊ ᑐࡋ࡚ࠊᙉไⓎ⌧ࢆ⾜ࡗࡓࡇࢁࠊᚓࡽࢀࡓᙧ㉁㌿ య࡚࡛ࡣ࡞࠸ࡀࠊѸᆺ⣽⬊ࡢᛶ㌿ᡂຌࡋࡓࠋ ࡇࡢ㑇ఏᏊࢆྵࡴѸᆺ VFDIIROG ࡣ NE ࡛࠶ࡗࡓ ࡀࠊヱᙜࡍࡿ VFDIIROG ࡣ㸩ᆺ⣽⬊ࡽぢ࠸ࡔࡉࢀ࡚ - 396 - ࠾ࡽࡎࠊѸᆺࢆỴᐃ࡙ࡅࡿᛶᰁⰍయᵝ㡿ᇦ࡛࠶ࡿྍ⬟ ᛶࡀ㧗࠸ࠋ ' 3& ࢣ࣑ࢫࢺ࣮ࣜࢆ⏝࠸ࡓ 3DF%LR56,, ࡼ ࡿࢤࣀ࣒ࢩ࣮ࢡ࢚ࣥࢫ ᅜ❧㑇ఏᏛ◊✲ᡤ࣭㇏⏣ඛ⏕ࡢࡶࠊ㸩ᆺ࠾ࡼ ࡧѸᆺࡑࢀࡒࢀࡢࢤࣀ࣒㓄ิࢆࠊ3& ࢣ࣑ࢫࢺ࣮ࣜ ࢆ⏝࠸ࡓ 3DF%LR56,, ࡚ྲྀᚓࡋࡓࠋࡑࢀࡒࢀ ࢭ ࣝࡎࡘྲྀᚓࡋ࡚࠾ࡾࠊ0E ࡋ࡚ [ ࡢ GHSWK ࡞ࡿணᐃ࡛࠶ࡿࠋ (3DF%LR ࢹ࣮ࢱཬࡧ ,OOXPLQD ࢹ࣮ࢱࡢࣁࣈࣜ ࢵࢻࢭࣥࣈࣜࢆ⾜࠺ࡓࡵࡢࠊゎᯒࣝࢦࣜࢬ࣒タ ィ ,OOXPLQDVKRUWUHDG ࢹ࣮ࢱࠊ3DF%LR ࢹ࣮ࢱ ࢆ⤌ࡳྜࢃࡏ࡚ࠊᴟຊ࢚࣮ࣛࢆ࢟ࣕࣥࢭࣝࡋࡓ⢭ ᗘࡢ㧗࠸ࢤࣀ࣒㓄ิࢆྲྀᚓࡍࡿࡇࡋࡓࠋࡑࡢ ࡓࡵࠊᮾ᪂㡿ᇦ࣭➟ཎ༤ኈࡢᨭࢆཷࡅࠊࣄ࣓࣑ ࢝ࢷ࢟ࣔࡢࢤࣀ࣒≉ᛶྜࢃࡏࡓᑓ⏝ࡢゎᯒࣝࢦ ࣜࢬ࣒ࡢタィࢆ㐍ࡵࡓࠋ ) 㓄ᏊᏞᰴ㸦 ᰴ㸧ࡢ࣑ࣝࢼ LQGH[ ࢩ࣮ࢣࣥ ࢫ 㓄ᏊᏞᰴ㸩ᆺ ⣔⤫ࠊ࣐ࢼࢫᆺ ⣔⤫ࡘ ࠸࡚ ,OOXPLQD,QGH[ࢩ࣮ࢣࣥࢩࣥࢢࢆ⾜࠸ࠊ㡿ᇦࢆ 㐃㙐⩌ศ㞳ࡍࡿࡇࡼࡗ࡚ࢭࣥࣈ࣮ࣜࢆᨭ ࡍࡿࢹ࣮ࢱࡋࡓࠋࡇࡢ⤖ᯝࢆࡶࠊᛶ⾲⌧㐃 㙐ࡍࡿྍ⬟ᛶࡢ࠶ࡿ㡿ᇦࢆ⤠ࡾ㎸ࡳࠊྠᐃࡋࡓ VFDIIROG ≉␗ⓗ 3&5 ࣉ࣐࣮ࣛࢆタィࡋࠊᑐᛂࡍࡿ %$& ࢡ࣮ࣟࣥࢆࢫࢡ࣮ࣜࢽࣥࢢࡍࡿணᐃ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᴫせࢤࣀ࣒㓄ิࡢ୰ࡽࠊࡇࢀࡲ࡛㝣ୖ᳜≀ࠊࡶ ࡋࡃࡣከ⣽⬊ࡢࢩࣕࢪࢡࣔࡲ࡛࡛▱ࡽࢀ࡚࠸ࡓ࠸ࡃ ࡘࡢ㌿ᅉᏊ⩌㸦ࢡࣛࢫ㸰.12; 㑇ఏᏊࠊ+'=,3 㑇ఏᏊ࡞㸧ࡀⓎぢࡉࢀࡓࠋࡲࡓࠊࢡ࣑ࣛࢻࣔࢼࢫ ࡞ࡢ⥳⸴ࡣᏑᅾࡏࡎࠊ㝣ୖ᳜≀Ꮡᅾࡍࡿ㌿ ᅉᏊࣇ࣑࣮ࣜ㸦 ✀㢮㸧ࡢ࠺ࡕࠊ ✀㢮ࡀࣄ࣓࣑ ࢝ࢷ࢟ࣔࡢࢤࣀ࣒୰Ꮡᅾࡋ࡚࠸ࡿࡇࡀุ᫂ࡋࡓࠋ ୍᪉ࠊᰤ㣴⏕Ṫᮇࠊ⏕Ṫ⏕㛗ᮇࡢษ㛵ࢃࡿ㌿ ᅉᏊ࡛࠶ࡿ /HDI\ ࡘ࠸࡚ࡣࠊᏑᅾࡀぢࡽࢀ࡞ࡗ ࡓࠋ㝣ୖ᳜≀ࡢከ⣽⬊ࠊከ⣽⬊⏕Ṫჾᐁᙧᡂ࡞ ࠾࠸࡚ࠊࢤࣀ࣒࡛ࣞ࣋ࣝࡢࡼ࠺࡞㐍ࡀ㉳ࡇࡗ ࡚ࡁࡓࡢࡘ࠸࡚ࠊᐇయゎ᫂ࡀ㐍ࡴࡇࡀᮇᚅࡉ ࢀࡓࠋ ୧᥋ྜᆺ⣽⬊ࡢࢤࣀ࣒ࢆẚ㍑ࡋࠊࡉࡽࠊ WUDQVFULSWRPH ࢹ࣮ࢱࡢ⤖ᯝࢆᴫせࢤࣀ࣒ࢹ࣮ࢱ ↷ࡽࡋྜࢃࡏࡿࡇ࡛ࠊѸᆺ⣽⬊ࢤࣀ࣒ࡢࡳᏑᅾࡋࠊ ぶᰴ࠺ࡋࡢࡅྜࢃࡏࡼࡾᚓࡽࢀࡓᏊᏞᰴࡢᛶ ⾲⌧㸦Ѹᆺ࡛࠶ࡿࡇ㸧㐃㙐ࡍࡿ &S0LQXV 㑇ఏᏊࡢⓎぢ⮳ࡗࡓࠋ&S0LQXV 㑇ఏᏊࡣࠊE=,3 ᆺ ࡢ㌿ᅉᏊࢆࢥ࣮ࢻࡋ࡚࠾ࡾࠊࡉࡽࡇࡢ㑇ఏᏊࢆ 㸩ᆺ⣽⬊࡛ᙉไⓎ⌧ࡋࡓࡇࢁࠊ㸩ᆺࡽѸᆺᛶ ㌿ࡋࠊ㸩ᆺ⣽⬊ࡢΰྜࡼࡾ᥋ྜࡍࡿࡇࡀ᫂ ࡽ࡞ࡗࡓࠋࡋࡋ࡞ࡀࡽࠊ㓄┦ᡭΰࡐࡎ ᙧ㉁㌿యࢆ༢⊂࡛᥋ྜㄏᑟ᮲௳࠾࠸ࡓሙྜࠊᑡ ᩘ࡛ࡣ࠶ࡿࡀࠊணࡋ࡚ࠊࢡ࣮ࣟࣥෆ࡛࣌ࢆ ࡘࡃࡾ᥋ྜᏊࢆᙧᡂࡍࡿࡇࡀุ᫂ࡋࡓࠋࡇࡢᐇ ࡣࠊձ&S0LQXV 㑇ఏᏊ⏘≀ࡼࡾѸᆺ⣽⬊ࡋ࡚ࡢᛶ ㉁ࡀⓎࡍࡿࡼ࠺࡞ࡗࡓࡇࠊղᙧ㉁㌿యࡣࠊ ౫↛ࡋ࡚㸩ᆺ⣽⬊ࡋ࡚ࡢᛶ㉁ࡶࢃࡎಖᣢࡋ ࡚࠸ࡿࡇࠊࢆ♧၀ࡋ࡚࠸ࡿࠋࡲࡓࠊࡇࡢ⤖ᯝࡣࠊ 㸩ᆺ⣽⬊ࡋ࡚ࡢࡿ⯙࠸㛵ࢃࡿᮍ▱㑇ఏᏊ &S3OXVࠊࡲࡓࡣࡑࡢࡿ⯙࠸ࢆᢚไࡍࡿᮍ▱㑇ఏᏊ &S0LQXV ࡢ ࠊ ࠸ ࡎ ࢀ ࠊ ࡲ ࡓ ࡣ ᪉ ࡀ Ꮡ ᅾ ࡋ ࠊ &S0LQXV ࢭࢵࢺ࡞ࡾࠊ࣊ࢸࣟᰴࡢᛶ⾲⌧ࢆỴᐃ ࡍࡿࡇࡶ♧၀ࡋ࡚࠾ࡾࠊࡇࡢ㛵ಀࡀᔂࢀࡿࡇ࡛ࠊ ࢡ࣮ࣟࣥෆ࡛᥋ྜᏊࢆࡘࡃࡿ࣍ࣔࢱࣜࢵࢡᰴ㸦࣍ࣔ ᰴ㸧ࡍࡿྍ⬟ᛶࡶᐃࡉࢀࡓࠋ ୍᪉ࠊࣄ࣓࣑࢝ࢷ࢟ࣔࡢ࣊ࢸࣟᰴࡣࠊࡇࢀࡲ࡛ ⏝࠸࡚ࡁࡓᇶ‽ᰴ㸦㓄⩌ ,(㸧ຍ࠼࡚ࠊ⏕Ṫ㝸㞳 ࡋࡓᑡ࡞ࡃࡶ㸱ࡘࡢ㓄⩌㸦,,$,,%,,&㸧ࡀᏑ ᅾࡋ࡚࠸ࡿࠋࡲࡓࠊࡑࢀࡒࢀ㏆⦕࡞࣍ࣔᰴࡀⓎぢ ࡉࢀࠊ㓄⩌ ,,% ᒓࡍࡿ࣊ࢸࣟᰴࡣࠊ㏆⦕࡞࣍ࣔ ᰴ㸦NRGDPD ᰴ㸧㒊ศⓗ࡞ᐙ᥋ྜᛂࢆ♧ࡍࠋ ࣍ࣔᰴࡢ 51$VHT ゎᯒࡢ⤖ᯝࠊ&S0LQXV ࢜ࣝࢯࣟࢢ㑇 ఏᏊࡢⓎぢ⮳ࡗࡓࠋࡇࡢ㑇ఏᏊࡢⓎ⌧ࢆᢚไࡋࡓ ᙧ㉁㌿య࡛ࡣࠊ⮬ᐙ᥋ྜ⬟ຊࡀపୗࡋࠊ㏆⦕࡞࣊ ࢸࣟᰴࡢΰྜࡼࡾࠊᐙ᥋ྜࡍࡿഴྥࡀ㧗ࡲࡗ ࡓࡇࡽࠊࣄ࣓࣑࢝ࢷ࢟ࣔ࠾ࡅࡿṪࠊ⮬Ṫ ࠸ࡗࡓ⏕ṪᵝᘧࡀࠊᛶỴᐃ㑇ఏᏊ⩌ࡢാࡁࡼࡾࠊ ẚ㍑ⓗᐜ᫆㌿ࡋ࠺ࡿࡇࡀ♧၀ࡉࢀࡓࠋ 㸺ᨭᡂᯝබ⾲ㄽᩥࣜࢫࢺ㸼 ࡞ࡋ - 397 - බເᨭㄢ㢟㸸2013~15 ᖺᗘ ࢮࣈࣛࣇࢵࢩࣗࡢ㦵᱁➽ᑐࡍࡿᚤ㔜ຊ⎔ቃࡢᙳ㡪 ᇶ┙◊✲㸦㹁㸧 Pathway ゎᯒࡼࡿᚤ㔜ຊ⎔ቃୗ➽ⴎ⦰࣓࢝ࢽࢬ࣒ࡢゎ᫂ཎᅉ⒪ἲࡢ㛤Ⓨ ᇶ┙◊✲㸦㹁㸧 ℩ཎ ῟Ꮚ࣭ி㒔Ꮫ⏕་⛉Ꮫ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ேయࡣᚤ㔜ຊ⎔ቃ࠾ࢀࡿࠊࡑࡢᛂࡋ࡚ ➽ⴎ⦰ࢆ㉳ࡇࡍࠋࡘࡲࡾࠊᆅୖ࠾࠸࡚ࡣ㔜ຊ㈇Ⲵ ࢆཷࡅࡿࡇࡼࡾ➽⫗ࢆ⥔ᣢࡍࡿᶵᵓࡀാ࠸࡚࠸ ࡿࡢ࡛࠶ࡿࠋࢮࣈࣛࣇࢵࢩࣗࡢᏱᐂᅾᐇ㦂ࡼ ࡾᚤ㔜ຊ⎔ቃࢆ㉳ᅉࡍࡿ➽ⴎ⦰࠾ࡼࡧ㐠ືᢚไ࣭ ຍ㱋ࢆ㉳ᅉࡍࡿ➽ⴎ⦰ࡢศᏊᇶ┙ࢆࢺࣛࣥࢫࢡࣜ ࣉࢺ࣮࣒ゎᯒ3DWKZD\ ゎᯒࢆ୰ᚰẚ㍑ࡋࠊࡑࡢඹ ㏻Ⅼ࠾ࡼࡧ┦㐪Ⅼࢆྠᐃࡍࡿࡇࡼࡾ➽ⴎ⦰➽ ⫗⥔ᣢ࣓࢝ࢽࢬ࣒ࢆ᫂ࡽࡍࡿࠋ ࡉࡽࡣࠊྛ➽ⴎ⦰᮲௳ୗ࡛Ⓨ⌧ኚືࡀㄆࡵࡽࢀ ࡿ㑇ఏᏊ⩌ࢆᣦᶆࠊᙉไ㐠ື➼ࡢᑐ⒪ἲ࡛ࡣ࡞ ࡃཎᅉ⒪ἲࡢ㛤Ⓨࢆヨࡳࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࢮࣈࣛࣇࢵࢩࣗࡢ㐠ືᢚไࠊຍ㱋ࠊᏱᐂᅾ㸦㌶ 㐨ୖ ᪥ᚋࠊ㌶㐨ୖ⣙㸯࢝᭶ᚋࠊᆅୖᖐ㑏ᚋ ᪥ᚋࠊ ᆅୖᖐ㑏ᚋ⣙㸯࢝᭶ᚋ㸧᮲௳ୗ࠾ࡅࡿ㦵᱁➽ࢺࣛ ࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࠋ ࡲࡓࠊཎᅉ⒪ἲࡢᑐ㇟ೃ⿵ศᏊ࡛࠶ࡿ P\RVWDWLQ 㐣Ⓨ⌧࠾ࡼࡧάᛶ㜼ᐖ᮲௳ୗ࠾ࡅࡿ㦵᱁➽ࢺࣛ ࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㐠ືᢚไ࠾ࡼࡧຍ㱋࠾࠸࡚ከᩘࡢ㑇ఏᏊⓎ⌧ 㔞ࡢኚࡀ☜ㄆࡉࢀࡓࠋ㐠ືᢚไࡼࡿ➽ⴎ⦰ࡣᗫ ⏝ᛶ➽ⴎ⦰⪃࠼ࡽࢀ࡚࠾ࡾࠊࣉࣟࢸࢯ࣮࣒⣔ࡢ άᛶࡀ᪤▱ࡽࢀ࡚࠸ࡿࠋᅇࡢࢮࣈࣛࣇࢵࢩ ࣗ㐠ືᢚไ࠾࠸࡚ࡶከᩘࡢࣉࣟࢸࢯ࣮࣒㛵㐃㑇 ఏᏊࡢⓎ⌧ቑຍࡀㄆࡵࡽࢀࡓࠋࡇࡢࡇࡣࢮࣈࣛࣇ ࢵࢩࣗ࠾࠸࡚ࡶ㐠ືᢚไࡼࡗ࡚ᗫ⏝ᛶ➽ⴎ⦰ ࡀㄏⓎࡉࢀ࡚࠸ࡿࡇࢆ♧၀ࡋ࡚࠾ࡾࠊࢮࣈࣛࣇ ࢵࢩࣗࡀ➽ⴎ⦰◊✲ࡢࣔࢹࣝື≀ࡋ࡚᭷⏝࡛࠶ࡿ ࡇࢆ♧ࡍ⤖ᯝ࡛࠶ࡿࠋࡲࡓࠊᚤ㔜ຊ⎔ቃᅾࡼ ࡿ➽ⴎ⦰ࡀᗫ⏝ᛶ➽ⴎ⦰㠀ᖖ㢮ఝࡋ࡚࠸ࡿ⪃ ࠼ࡽࢀ࡚࠸ࡿࡇࡽࡶࠊࢮࣈࣛࣇࢵࢩ࡛ࣗᗫ⏝ ᛶ➽ⴎ⦰ࡢഴྥࡀ☜ㄆࡉࢀࡓࡇࡣ㠀ᖖ⯆῝࠸ ⤖ᯝ࡛࠶ࡿࠋ ୍᪉ࠊຍ㱋㛵ࡋ࡚ࡣ㐠ືᢚไྠᵝ࠸ࡃࡘ ࡢࣉࣟࢸࢯ࣮࣒㛵㐃㑇ఏᏊࡢⓎ⌧ቑຍࡀㄆࡵࡽࢀ ࡓࡀࠊ㐠ືᢚไ࡛ࡣኚࡀㄆࡵࡽࢀ࡞ࡗࡓ㑇ఏᏊ 㛵ࡋ࡚ࡶከᩘⓎ⌧㔞ࡀኚࡋࡓ㑇ఏᏊࡀ☜ㄆࡉࢀ ࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡢ୰ࡣ➽ⴎ⦰ᛶᝈࡢཎᅉ㑇 ఏᏊࡶྵࡲࢀ࡚࠾ࡾࠊ᪂つࡢຍ㱋ࡼࡿ➽ⴎ⦰࣓࢝ ࢽࢬ࣒ࡢゎ᫂ᙺ❧ࡘ⤖ᯝ࡛࠶ࡿࠋ Ᏹᐂᅾᐇ㦂㛵ࡋ࡚ࡣࠊ㌶㐨ୖ ᪥ᚋࠊ㌶㐨ୖ ⣙㸯࢝᭶ᚋࠊึࡵ࡚ᡂຌࡋࡓᆅୖᖐ㑏ᚋࡢࢮࣈࣛࣇ ࢵࢩࣗࢆヨᩱࡋ࡚⏝࠸ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎ ᯒࢆ⾜ࡗࡓࠋ㌶㐨ୖ⣙㸯࢝᭶ᚋࡢゎᯒ⤖ᯝࡼࡾࠊࡇ ࢀࡲ࡛ࡢሗ࿌ྠᵝࡢⓎ⌧ኚືࢆ♧ࡍ㑇ఏᏊࡶ☜ㄆ ࡉࢀࡓࡀࠊ㏫ࡢኚືࢆ♧ࡍ㑇ఏᏊࡶከᩘ☜ㄆࡉࢀࠊ 㠀ᖖ⯆῝࠸⤖ᯝࡀᚓࡽࢀࡓࠋࡲࡓࠊ⤒ⓗ㸦ᆅ ୖȝ㌶㐨ୖȝᆅୖ㸧࡞㑇ఏᏊⓎ⌧ኚືࢆゎᯒࡍࡿࡇ ࡼࡾࠊᏱᐂᅾ࠾ࡼࡧᆅ⌫ᖐ㑏ᑐࡋ࡚᪩ᮇⓎ ⌧㔞ࡀኚࡍࡿ㑇ఏᏊࢆྠᐃࡍࡿࡇࡀ࡛ࡁࡓࠋࡇ ࢀࡽࡢ㑇ఏᏊࡢ୰ࡣࠊ᪤➽ⴎ⦰ࡢ㛵㐃ࡀሗ࿌ ࡉࢀ࡚࠸ࡿ㑇ఏᏊࡶྵࡲࢀ࡚࠾ࡾࠊヲ⣽࡞ゎᯒࢆ㐍 ࡵࡿ࠶ࡓࡾᑐ㇟ࡍࡿᅉᏊࡢ㑅ᐃ㠀ᖖ᭷⏝࡞ ࢹ࣮ࢱࡀᚓࡽࢀࡓࠋ ࡲࡓࠊᏱᐂᅾࡼࡿ➽ⴎ⦰ࡢཎᅉ⒪ἲࡢ㛤Ⓨࢆ ┠ⓗࡋ࡚ೃ⿵ᅉᏊ࡛࠶ࡿ P\RVWDWLQ ࡢ㐣Ⓨ⌧࠾ ࡼࡧάᛶ㜼ᐖ᮲௳ୗ࠾ࡅࡿ㦵᱁➽ࢺࣛࣥࢫࢡࣜࣉ ࢺ࣮࣒ゎᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊᏱᐂᅾࡼࡾⓎ ⌧ኚືࡀ☜ㄆࡉࢀࡓ㑇ఏᏊࡢᙳ㡪ࡀ☜ㄆࡉࢀࠊᏱ ᐂᅾࡼࡿ➽ⴎ⦰ࡢཎᅉ⒪ἲࡋ࡚ P\RVWDWLQ ά ᛶᢚไࡢྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ ࡇࢀࡽࡢ⤖ᯝࡣᏱᐂ✵㛫ᅾ࠾ࡅࡿ➽ⴎ⦰࣓࢝ ࢽࢬ࣒ࡸཎᅉ⒪ἲࡢ㛤Ⓨࡔࡅ࡛࡞ࡃࠊ㧗㱋⪅ࡸ㛗ᮇ ධ㝔ᝈ⪅࠾ࡅࡿ㦵᱁➽⥔ᣢ࣭ᅇ࡞ࠊண㜵ࡸ ⒪ἲࡢ㛤Ⓨࡶ㈉⊩ࡍࡿ⪃࠼ࡽࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢮࣈࣛࣇࢵࢩࣗࡢ➽ⴎ⦰◊✲ࡢࣔࢹࣝື≀ࡋ ࡚ࡢ᭷⏝ᛶ࠾ࡼࡧᏱᐂᅾ᮲௳ୗࡢ➽ⴎ⦰◊✲࠾ ࡅࡿࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࡢ᭷ຠᛶࢆ☜ㄆࡍࡿ ࡇࡀ࡛ࡁࡓࠋ⌧ᅾᏱᐂᐇ㦂࠾࠸࡚Ᏻᐃⓗࡘ㛗 ᮇⓗ⏝࠸ࡽࢀ࡚࠸ࡿ⬨᳝ື≀ࡣỈ᳇⏕≀㸦ࢮࣈࣛ ࣇࢵࢩࣗࡸ࣓ࢲ࢝➼㸧㝈ࡽࢀ࡚࠾ࡾࠊỈ᳇ᐇ㦂 ⨨ࢆഛࡍࡿࡢࡣୡ⏺࡛ࡶ᪥ᮏࡔࡅ࡛࠶ࡿࠋࡇࡢ ࡇࡽࡶୡ⏺ⓗᗈࡃᐇ㦂ື≀ࡋ࡚⏝ࡉࢀ࡚ ࠸ࡿࢮࣈࣛࣇࢵࢩࣗࢆ⏝࠸ࡓ➽ⴎ⦰◊✲ࡢྍ⬟ᛶ ࢆ☜ㄆ࡛ࡁࡓࡇࡼࡾࠊᚋࡢᏱᐂᐇ㦂ࡢࡳ࡞ࡽ ࡎ➽ⴎ⦰ᛶᝈ➼ࡢ◊✲࠾࠸࡚ࡶ᪂ࡓ࡞➽ⴎ⦰◊ ✲ࡢศ㔝ࢆ㛤ᣅࡍࡿࡇࡀ࡛ࡁࡓࠋࡲࡓࠊ᪂ࡓᅜ 㝿Ᏹᐂࢫࢸ࣮ࢩࣙࣥ㸦ISS㸧ࡢࠕࡁࡰ࠺ࠖ᪥ᮏᐇ㦂 Ჷࡢ⯪ෆ⎔ቃࢆ⏝ࡍࡿᐇ㦂ࢸ࣮࣐㸦ࣇࢪࣅࣜ ࢸࢫࢱࢹࢸ࣮࣐㸧㑅ᐃࡉࢀࠊࡼࡾヲ⣽࡞ゎ ᯒࢆ┠ⓗࡋࡓᏱᐂᅾᐇ㦂ࢆ⾜࠺ᶵࢆᚓࡿࡇ ࡀ࡛ࡁࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Ⓨ⾲‽ഛ୰ࠋ - 398 - බເᨭㄢ㢟㸸2013 ᖺᗘ ࣐࣐࢝ࣈࣜࡢࢤࣀ࣒ሷᇶ㓄ิ㐺ᛂᙧែ㑇ఏᏊ ᪂Ꮫ⾡㡿ᇦ◊✲ ᭮⏣ ㈆࣭ி㒔ᏛᏛ㝔⌮Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ≉ᐃࡢ㣵ື≀ࡢᑓ㛛ࡀ㸪ࡦࡘࡢ⣔⤫⩌ࡢᙧ ែⓗከᵝ✀ࡢከᵝ㸪ࡍ࡞ࢃࡕ㐺ᛂᨺᩓࢆࡶࡓ ࡽࡍሙྜࡀ࠶ࡿ㸬ୡ⏺ࡢ ᖏᇦࡑࡢ࿘㎶ศᕸࡍ ࡿᆅ⾲ᛶᤕ㣗⪅࢜ࢧ࣒ࢩ㢮 1000 ✀௨ୖࡢ࠺ࡕ㸪⣙༙ ᩘࡣ࢝ࢱࢶ࣒ࣜ㸦㝣㈅㸧㣗ᑓ㛛ࡋ࡚࠸ࡿ㸬ࡇࢀ ࡽࡢ㈅㣗ᛶ࢜ࢧ࣒ࢩࡣ㸪ᤕ㣗⾜ືࡢ㐪࠸㛵㐃ࡋ࡚㸪 ᕧ㢌ᆺ⊈㢌ᆺࡢศࢆ♧ࡍ㸬ᕧ㢌ᆺࡣ㸪ẚ㍑ⓗᑠ ࡉ࠸㝣㈅ࡢẆࢆჶࡳ◚ࡗ࡚㌾య㒊ࢆ㣗㸪⊈㢌ᆺࡣ ẚ㍑ⓗࡁ࡞㝣㈅ࡢẆࡢ୰㢌㒊ࢆ✺ࡗ㎸ࢇ࡛㌾య 㒊ࢆ㣗ࡿ㸬ᮏ◊✲࡛ࡣ㸪᪥ᮏᅛ᭷ࡢ㈅㣗ᛶ࢜ࢧ࣒ ࢩ㸪࣐࣐࢝ࣈࣜ Damaster blaptoides ࡛ࡣள✀㛫 ࡛ࡇࡢᙧែศࡀ㉳ࡇࡗ࡚࠾ࡾ㸪ࢤࣀ࣒ࢧࢬࡀẚ ㍑ⓗᑠࡉࡃ㸪ࢤࣀ࣒ゎㄞࢆヨࡳ࡞ࡀࡽ㸪㐺ᛂⓗᙧែ ศࡢ㑇ఏᏊࢆゎ᫂ࡍࡿୖ࡛᭷⏝࡞◊✲ᑐ㇟࡛࠶ࡿ㸬 ᮏ◊✲࡛ࡣ㸪࣐࣐࢝ࣈࣜࡢࢤࣀ࣒ࢆゎㄞࡍࡿ ྠ㸪ள✀㛫ࡢᙧែᕪ㛵ࡍࡿ㧗⢭ᗘࡢ QTL ࣐ࢵ ࣆࣥࢢࢆ⾜࠺㸬ࡲࡓ௦⾲ⓗ࡞ள✀ࡘ࠸࡚㸪ࢺࣛࣥ ࢫࢡࣜࣉࢺ࣮࣒ẚ㍑㸪ࢤࣀ࣒ሷᇶ㓄ิࡢࣜࢩ࣮ࢣࣥ ࢩࣥࢢࢆ⾜࠸㸪㐺ᛂᙧែ㐍ࡢ㑇ఏⓗᇶ┙ࢆゎ᫂ࡍ ࡿ㸬 㸺ᨭࡢෆᐜ㸼 ࣐࣐࢝ࣈࣜ⢖ᓥள✀㸦࣐࣐࢜࢝ࣈࣜ D. b. fortunei㸧࣐࣐࢝ࣈࣜࡢࢤࣀ࣒㓄ิゎㄞ ⏝ࡍࡿ㸪Nextera Matepair Kit ࡼࡿ NE ࣥࢧ࣮ ࢺࡢ࣓ࢺ࣌ࢩ࣮ࢣࣥࢫࢹ࣮ࢱࡢྲྀᚓ㸦ᙜึࡣ fosmid ࡼࡿࣥࢧ࣮ࢺ㛗 40kb ࡢ࣓ࢺ࣌ࢩ࣮ ࢣࣥࢫࡀ⾜ࢃࢀࡿணᐃ࡛࠶ࡗࡓࡀ㸪౫㢗ࡋࡓࢧࣥࣉ ࡛ࣝࡣᵝࠎ࡞᮲௳࡛ヨࡳ࡚ࡶ fosmid ࣛࣈ࣮ࣛࣜࡢ ࢱࢱ࣮ࡀ༑ศ㔞㐩ࡏࡎ㸪Nextera Matepair Kit ࡼࡿ 20kb ࢩ࣮ࢣࣥࢫኚ᭦ࡉࢀࡓ㸬㸧 Ꮚࡢ᥈⣴ࡢࡓࡵ㸪RAD ࢩ࣮ࢣࣥࢫࢆ⏝࠸ࡓ㐃㙐ᆅ ᅗసᡂᙧែᙧ㉁ࡢ QTL ゎᯒ㸪ྛⓎ⫱ࢫࢸ࣮ࢪ࠾ ࡅࡿள✀㛫ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ẚ㍑ゎᯒࢆ⾜ࡗ ࡓ㸬RAD-seq ἲࡼࡗ࡚⋓ᚓࡋࡓศᏊ࣐࣮࣮࢝ ࢆ࠸㐃㙐ゎᯒࢆ⾜ࡗࡓ⤖ᯝ㸪ᰁⰍయྠᩘࡢ 㐃㙐⩌ࡽᵓᡂࡉࢀࡿ㧗ᐦᗘ㐃㙐ᆅᅗࢆస〇ࡍ ࡿࡇࡀ࡛ࡁࡓ㸬ࡇࡢ㐃㙐ᆅᅗࢆ⏝࠸࡚ QTL ࣐ ࢵࣆࣥࢢࢆ⾜ࡗࡓ⤖ᯝ㸪࣐࣐࢝ࣈࣜእ㒊ᙧ ែ㛵ࢃࡿ QTL 㡿ᇦࢆ≉ᐃࡍࡿࡇࡀ࡛ࡁࡓ㸬 ᚋࡣࢭࣥࣈࣝ⤖ᯝᇶ࡙࠸ࡓࣀࢸ࣮ࢩࣙ ࣥሗ㑇ఏᏊⓎ⌧ゎᯒ⤖ᯝࢆࡶ㸪ࡇࡢ QTL 㡿ᇦࡢ㑇ఏᏊ≉ᐃࢆ㐍ࡵࡿ㸬௨ୖࡢ⤖ᯝࡣ㸪 ࢜ࢧ࣒ࢩ㢮ࡢ㐺ᛂ㐍ࢆࢤࣀ࣒࡛ࣞ࣋ࣝ㆟ㄽࡍ ࡿୖ࡛㠀ᖖព⩏῝࠸⤖ᯝ࡛࠶ࡿࡶ㸪 ᚋ㸪᪻ᙧែࡢከᵝࢆẚ㍑ࢤࣀ࣒ⓗ࡞ࣉࣟ ࣮ࢳ࡛ゎ᫂ࡍࡿୖ࡛㔜せ࡞◊✲⤖ᯝ࡞ࡿࡇ ࡀᮇᚅ࡛ࡁࡿ㸬 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᭮⏣㈆࣭ᑠ㡰࣭⸨⃝▱ぶ㸦㸧㝣㈅㣗࢜ ࢧ࣒ࢩࡢᕧ㢌ᆺ⊃㢌ᆺ㸸㐺ᛂⓗᙧែศࡣࡢ ࡼ࠺㉳ࡇࡿࡢ㸽 ࢤࣀ࣒࡛㐍ࡢㅦࢆゎࡃ㸟 ➨ ᅇ㸬⣽⬊ᕤᏛ 㸦ᰝㄞ↓㸧 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣥࢧ࣮ࢺ㛗 180 bp㸪500 bp ࡢ࢚࣌ࣥࢻࢩ࣮ࢣ ࣥࢫ㸪ᮏᨭ࡛ᚓࡽࢀࡓࣥࢧ࣮ࢺ㛗 20 kb ࡢ࣓ ࢺ࣌ࢩ࣮ࢣࣥࢫࡇࡢᨭࡣูᚓࡓࣥࢧ࣮ ࢺ㛗 2.5 kb, 3.5 kb, 4.5 kb, 7 kb, 11 kb ࡢ࣓ࢺ࣌ ࢩ࣮ࢣࣥࢫࡢࢹ࣮ࢱࢆ࠸㸪Platanus 1.2.1 ࡼࡿ ࢭࣥࣈࣝࢫ࢟ࣕࣇ࢛ࣝࢹࣥࢢࢆ⾜ࡗࡓ㸬ࡑࡢ ⤖ᯝ㸪㛗⣙ 180 Mb㸪scaffold N50 ࡀ⣙ 3.5 Mb ࡢ ࢤࣀ࣒㓄ิࡀࢭࣥࣈࣝࡉࢀࡓ㸬ࡲࡓ㸪PacBioRS ࡼࡗ࡚ᖹᆒ᭱ࢧࣈ࣮ࣜࢻ㛗 1,654bp㸪ࢤࣀ࣒ࢧ ࢬ 40 ಸࡢࢩ࣮ࢣࣥࢫࢆྲྀᚓࡋ࡚࠾ࡾ㸪ᚓࡽࢀࡓࢫ ࢟ࣕࢵࣇ࢛ࣝࢻᑐࡋ PBjelly 13.10.22 ࡼࡿࢠࣕ ࢵࣉࢡ࣮ࣟࢪࣥࢢࢆ⾜ࡗࡓ⤖ᯝࠊcontig N50 ࢆᨵၿ ࡉࡏࡿࡇࡀ࡛ࡁࡓ㸬 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲࡛ࡣ㸪ࢤࣀ࣒ゎㄞຍ࠼࡚㸪࣐࣐࢝ࣈ ࣜࡢ⊈㢌ᆺ࣭ᕧ㢌ᆺள✀㛫ࡢᙧែᕪ㛵ࡍࡿ㑇ఏ - 399 - බເᨭㄢ㢟㸸2010㹼2012 ᖺᗘ ㌿ไᚚࢿࢵࢺ࣮࣡ࢡࡽぢࡿཎཱྀᙧᡂཎ⭠⬇࣮࢜࢞ࢼࢨ࣮ࡢ㐍ࡢ࣓࢝ࢽࢬ࣒ ᇶ┙◊✲㸦㹀㸧 ᖹⰋ ┾つ࣭ᮾிᏛᏛ㝔⌮Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⬨᳝ື≀ึᮇⓎ⏕ࡢཎཱྀᙧᡂཎ⭠⬇࣮࢜࢞ࢼࢨ ࣮ࡣᇶᮏⓗ࣎ࢹࣉࣛࣥᙧᡂ࠾࠸࡚㔜せ࡞ᙺࢆ ᢸ࠺ࠋࡇࡢⓎ⏕ᵝᘧࡀ┿ṇᚋ⏕ື≀ࡢ㐍ࡢ㐣⛬࡛ ࡢࡼ࠺⋓ᚓࡉࢀࡓࢆ㑇ఏᏊ࡛ࣞ࣋ࣝゎᯒࡋࠊ ࡑࡢ㐍ࢩࢼࣜ࢜ࢆసᡂࡍࡿࡇࡣ㔜せ࡞◊✲ㄢ㢟 ࡛࠶ࡿࠋࡇࢀࡲ࡛ࡇࡢࡼ࠺࡞ㄢ㢟ࢆᐇドⓗゎᯒࡍ ࡿ᭷ຠ࡞ᡭẁࡀ࡞ࡗࡓࡀࠊ㏆ᖺࠊ✀ࠎࡢ⏕≀ࡢࢤ ࣀ࣒ࡢゎㄞචỿ㝆࣭ࢩ࣮ࢣࣥࢫ㸦ChIP-seq㸧ἲ ࡀ㛤Ⓨࡉࢀࡓࡇ࡛ࠊ⌧Ꮡࡢ┿ṇᚋ⏕ື≀ࡢ㌿ไ ᚚࢿࢵࢺ࣮࣡ࢡࢆ⥙⨶ⓗゎᯒࡍࡿࡇࡀྍ⬟࡞ ࡾࠊࡑࢀᇶ࡙࠸࡚㐍ࢩࢼࣜ࢜ࢆసᡂࡍࡿࡇࡀ ཎ⌮ⓗྍ⬟࡞ࡗࡓࠋࡑࡇ࡛ᮏ◊✲ࡣࠊࡲࡎ୧⏕ 㢮ࡢཎ⭠⬇ࢆ⏝࠸࡚ ChIP-seq ἲࡢ᭷ຠᛶࢆ᳨ドࡋ ࡘࡘࠊ࣮࢜࢞ࢼࢨ࣮࠾ࡅࡿ㌿ไᚚࢿࢵࢺ࣮࣡ ࢡࢆゎᯒࡍࡿࡇ࡛ࠊࡑࡢᇶ♏ࢆ☜❧ࡍࡿࡇࢆ┠ ᣦࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 Ⳣ㔝◊✲ᐊࡽᨭࢆཷࡅࠊࢿࢵࢱࢶ࣓࢚࢞ࣝ 㸦 Xenopus/Silurana tropicalis㸧ࡢཎ⭠⬇ࢆ⏝࠸ࡓ ChIP-seq ゎᯒཎ⭠⬇ࡢ RNA-seq ゎᯒ࠾ࡅࡿࠊ massive parallel sequencing ࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ ChIP-seq ゎᯒ࡛⏝࠸ࡓᢠయࡣࠊ6 ✀㢮ࡢ㌿ᅉᏊ Otx2ࠊLim1 (Lhx1)ࠊGsc (Goosecoid)ࠊMix1ࠊVegTࠊ Sia (Siamois)ࠊ㌿άᛶඹᙺᅉᏊࡢ p300ࠊ㌿ ᢚ ไ ඹ ᙺ ᅉ Ꮚ ࡢ TLE/Groucho ࠊ ࣄ ࢫ ࢺ ࣥ ಟ 㣭 ࡢ H3K4me1ࠊH3K27ac ࡛࠶ࡿࠋRNA-seq ゎᯒ࡛ࡣࠊ ཎ⭠⬇ࡢ⬇࡞ࡽࡧࠊཎ⭠⬇ࡽษࡾฟࡋࡓ㢌㒊 ࣮࢜࢞ࢼࢨ࣮㡿ᇦࠊ⫼ഃ㡿ᇦࠊ⭡ഃ㡿ᇦࠊᖏᇦ࡞ ࡽᢳฟࡋࡓ RNA ࢆ⏝࠸ࡓࠋChIP-seq ゎᯒ ࠾ࡅࡿࣆ࣮ࢡ᳨ฟࡣࠊX. tropicalis ࢤࣀ࣒㓄ิࢹ࣮ࢱ ࣮࣋ࢫ ver. 4㸦2012 ᖺࡲ࡛⏝㸧࠾ࡼࡧ ver. 7 ษ ࡾ᭰࠼ࡓᙜึࡣࠊⳢ㔝◊✲ᐊ࡛ࢃࢀ࡚࠸ࡓ⮬సࡢ ࣆ࣮ࢡ᳨ฟࢶ࣮ࣝࢆ⏝࠸ࡓゎᯒ࡛ᨭࢆཷࡅࡓࠋࡑ ࡢᚋࠊࣆ࣮ࢡ᳨ฟࡢᶆ‽ࢶ࣮ࣝࡉࢀࡿ MACS ࢆ⏝ ࠸࡚ᙜ◊✲ᐊ࡛⾜࠺ࡇࡋࠊࡑࡢࡓࡵࡢࢺ࣮ࣞࢽ ࣥࢢ࡛ᨭࢆཷࡅࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㸯㸧Otx2ࠊLim1ࠊGsc ࡢ ChIP-seq ゎᯒ࡞ࡽࡧ RNA-seq ゎᯒ Otx2ࠊLim1ࠊGsc ࡣ⬨᳝ື≀ࡢ㢌㒊࣮࢜࢞ࢼࢨ࣮ Ⓨ⌧ࡍࡿ࣓࣍࢜ࢻ࣓ࣥ㌿ᅉᏊ࡛࠶ࡿࠋࡇࢀࡽ ࡣ㢌㒊࣮࢜࢞ࢼࢨ࣮ࡢᙧᡂᶵ⬟㔜せ࡛࠶ࡿࡇ ࡀ 1990 ᖺ௦๓༙ࡼࡾㄪࡽࢀ࡚ࡁ࡚࠸ࡓࡀࠊࡢ ࡼ࠺࡞㑇ఏᏊ⩌ࢆไᚚࡋ࡚࠸ࡿࠊࡲࡓࡇࢀࡽ 3 ⪅ ࡣᶵ⬟ⓗࡢࡼ࠺㛵㐃ࡋ࡚࠸ࡿࡘ࠸࡚ࡣࠊ ᫂࡞Ⅼࡀᩘከࡃṧࡉࢀ࡚࠸ࡓࠋࡑࡇ࡛ Otx2ࠊLim1ࠊ Gsc ࡢ ChIP-seq ゎᯒࢆ⾜࠺ࡇ࡛ࠊࡑࢀࡽࡢᶆⓗ㑇 ఏᏊࢆ⥙⨶ⓗゎᯒࡋࠊࡉࡽࡑࢀࡽຍ࠼࡚ࠊ㌿ άᛶᢚไࢆ♧ࡍᣦᶆࡋ࡚ࠊ㌿άᛶඹᙺ ᅉᏊ p300ࠊ㌿ᢚไඹᙺᅉᏊ TLE/ࠊ࡞ࡽࡧάᛶ ᆺ ࣄ ࢫ ࢺ ࣥ ಟ 㣭 ࡢ H3K4me1 ࠊ H3K27ac ࡢ ChIP-seq ゎ ᯒࢆ ⾜ࡗࡓࠋᢚไᆺࣄࢫࢺࣥಟ㣭 ࡢ H3K27me3 ࡣሗ࿌ࡉࢀ࡚࠸ࡿࢹ࣮ࢱࢆ⏝࠸ࡓࠋ ࡇࢀࡽࡢࢹ࣮ࢱࢆᇶࠊࡲࡎ㑇ఏᏊࡢࢩࢫไᚚࣔࢪ ࣮ࣗࣝ㸦cis-regulatory module: CRM㸧ࡢྠᐃࢆ⾜ ࡗࡓࠋChIP-seq ࣆ࣮ࢡࡣഇࣆ࣮ࢡࡀྵࡲࢀࡿࡇ ࡣ☜⋡ㄽⓗணࡉࢀࠊࡲࡓ㌿ᅉᏊࡢ⤖ྜ㒊ࡀ ᚲࡎࡋࡶ CRM ࡣ㝈ࡽ࡞࠸ࡓࡵࠊ㌿ᅉᏊ㌿ ඹᙺᅉᏊࡢ࠺ࡕᑡ࡞ࡃࡶ 2 ࡘࡢࣆ࣮ࢡࡀ㔜࡞ࡗࡓ 㡿ᇦࢆ CRM ௬ᐃࡋ࡚ゎᯒࡍࡿࡇࡋࡓࠋ୍᪉ࠊ ไᚚࡉࢀࡿ㑇ఏᏊࡣࠊCRM ᭱ࡶ㏆ഐࡢ㑇ఏᏊࡋ ࡓࠋ Ⓨ⌧ࣃࢱ࣮ࣥࡼࡿ㑇ఏᏊࡢศ㢮ࡣࠊRNA-seq ࢹ࣮ ࢱࢆᇶ⾜࠸ࠊ㢌㒊࣮࢜࢞ࢼࢨ࣮≉␗ⓗ㑇ఏᏊ࡞ ࡀྵࡲࢀࡿ㡿ᇦ≉␗ⓗ㑇ఏᏊࠊ㡿ᇦ≉␗ᛶࡢ࡞ ࠸ࣘࣅ࢟ࢱࢫ㑇ఏᏊศ㢮ࡋࡓࠋࡇࢀࡽࢆ⏝࠸࡚ࠊ 㑇ఏᏊࡢ㡿ᇦ≉␗ᛶ p300 TLE ࡢ㛵ಀᛶࢆㄪ ࡓ⤖ᯝࠊ㡿ᇦ≉␗ᛶ㛵㐃ࡀ࠶ࡿᚑ᮶ሗ࿌ࡉࢀ࡚ ࠸ࡓ p300 ࡼࡾࡶࠊTLE ࡢ᪉ࡀࡼࡾᙉ࠸㛵㐃ᛶࢆࡶ ࡘࡇࠊ࠾ࡼࡧ㑇ఏᏊ࠶ࡓࡾࡢ TLE ⤖ྜ CRM ࡢᩘ ࡀከ࠸㡿ᇦ≉␗ᛶࡀᙉࡲࡿࡇࢆぢฟࡋࡓࠋࡘ ࡲࡾ㡿ᇦ≉␗ⓗ㑇ఏᏊࡣࠊⓎ⌧ࡋ࡚࠸࡞࠸㡿ᇦ࡛ TLE ࢆࡋ࡚Ⓨ⌧ࡀᢚไࡉࢀ࡚࠸ࡿࡇ࡞ࡽ࡞ ࠸ࠋᅇࡢゎᯒࡢࡼ࠺ࠊ⬇ࢆ⏝࠸ࡓ ChIP-seq ࢹ࣮ࢱࡣࠊⓎ⌧ࢆࡋ࡚࠸ࡿ⣽⬊ࡋ࡚࠸࡞࠸⣽⬊ࡀ ΰᅾࡍࡿࡓࡵࠊྠ୍ࡢ CRM p300 TLE ࡢࣆ࣮ ࢡࡀ㔜࡞ࡿࡇ࡞ࡿࠋOtx2 ࡣ㡿ᇦ≉␗ⓗ࡞㑇ఏᏊ ࢆไᚚࡍࡿ⪃࠼ࡽࢀࡿࡇࡽࠊOtx2 ࡀ⤖ྜࡍࡿ CRM ࡢ୰ࡽࠊTLE ࡶ⤖ྜࡋ࡚࠸ࡿ Otx2/TLE ⤖ ྜ CRM ὀ┠ࡋࡓࠋ Otx2/TLE ⤖ྜ CRM ࡣ㒊࡛ 13,443 ಶࡀྠᐃࡉࢀ ࡓࠋࡑࢀࡽ㏆ഐࡢ㑇ఏᏊࢆㄪࡓ⤖ᯝࠊணእ ࡶ㢌㒊࣮࢜࢞ࢼࢨ࣮㑇ఏᏊࡤࡾ࡛࡞ࡃࠊ⬗㒊࢜ ࣮࢞ࢼࢨ࣮ࡸ⭡ഃⓎ⌧ࡍࡿ㑇ఏᏊࡶྵࡲࢀ࡚࠸ ࡓࠋࡇࡢᐇࡽᡃࠎࡣࠊOtx2 ࡣ㢌㒊࣮࢜࢞ࢼࢨ ࣮㑇ఏᏊࢆάᛶࡍࡿ୍᪉࡛ࠊ㢌㒊࣮࢜࢞ࢼࢨ࣮ Ⓨ⌧ࡋ࡚࠸࡞࠸㑇ఏᏊࢆᢚไࡍࡿࡢ࡛ࡣ࡞࠸ ணࡋࡓࠋࡉࡽࠊࡇࡢάᛶࡢ㐪࠸ࢆࡶࡓࡽࡍࡢࡀࠊ ㌿άᛶᅉᏊࡋ࡚ᶵ⬟ࡍࡿ Lim1 ࠊ㌿ᢚไ ᅉᏊࡋ࡚ᶵ⬟ࡍࡿ Gsc ࡛ࡣ࡞࠸⪃࠼ࡓࠋࡘࡲ ࡾ Otx2 ࡣ Lim1 ඹ㑇ఏᏊࢆάᛶࡋࠊ ୍᪉ Otx2 ࡣ Gsc ඹ㑇ఏᏊࢆᢚไࡍࡿࠊ࠸࠺సᴗ௬ㄝࢆ ❧࡚ࡓࠋࡑࢀࢆ᳨ドࡍࡿࡓࡵࠊࡲࡎ Otx2/TLE/Lim1 ⤖ྜ CRM Otx2/TLE/Gsc ⤖ྜ CRM ࡢ 2 ⩌ศ ࡅࠊࡑࢀࡽ≉ᚩⓗ࡞㌿ᅉᏊ⤖ྜ㓄ิࢆ MIME ࣉ ࣟࢢ࣒ࣛࡼࡾゎᯒࡋࡓࡇࢁࠊ๓⪅ࡣ Lim1 ⤖ྜ 㓄ิࡀࠊᚋ⪅ࡣ Otx2/Gsc ⤖ྜ㓄ิࡀ⃰⦰ࡉࢀ࡚࠸ ࡓࠋࡑࡇ࡛ Otx2/TLE/Lim1 ⤖ྜ CRM ࡢ୰࡛ Lim1 ⤖ ྜ 㓄 ิ ࢆ ࡶ ࡘ ࡶ ࡢ ࢆ type I 㸦 1250 ಶ 㸧ࠊ Otx2/TLE/Gsc ⤖ྜ CRM ࡢ࠺ࡕ Otx2/Gsc ⤖ྜ㓄ิ ࢆࡶࡘࡶࡢࢆ type II㸦1039 ಶ㸧ࡋ࡚ࠊࡉࡽ᳨ウ - 400 - ࡋࡓࠋࡑࡢ⤖ᯝࠊtype I CRM ࡣ㢌㒊࣮࢜࢞ࢼࢨ࣮ 㑇ఏᏊࡢ㏆ഐࠊtype II CRM ࡀ㠀㢌㒊㑇ఏᏊࡢ㏆ ഐࡋ࡚Ꮡᅾࡋࡓࠋࡉࡽࣇࣜ࢝ࢶ࣓࢚࢞ࣝ ⬇ࡢணᐃእ⬇ⴥ㡿ᇦࢆ⏝࠸ࡓ࣏࣮ࣞࢱ࣮ゎᯒࡼࡾࠊ type I CRM ࡣࠊOtx2ࠊLim1ࠊGsc ࡢ 3 ⪅ࢆඹⓎ⌧ ࡉࡏࡓ≧ἣ㸦㢌㒊࣮࢜࢞ࢼࢨ࣮࢝ࢡࢸࣝ㸧࡛άᛶ ࡉࢀࡓ୍᪉ࠊtype II CRM ࡣྠࡌ㢌㒊࣮࢜࢞ࢼ ࢨ࣮࢝ࢡࢸ࡛ࣝᢚไࡉࢀࡓࠋࡇࢀࡽࡢ⤖ᯝࢆᇶࠊ ᡃࠎࡣ Otx2 ࡢᙺࡋ࡚௨ୗࡢࣔࢹࣝࢆᥦၐࡋࡓࠋ ࡍ࡞ࢃࡕࠊᩘ༓ࡢ㑇ఏᏊࡣࠊ㏆ഐࡢ CRM Otx2 ࡀ ⤖ྜࡍࡿࡇ࡛ࠊ⮬㌟ࡀ㢌㒊㡿ᇦ࠸ࡿㄆ㆑࡛ࡁࠊ ࡑࢀᇶ࡙ࡁ㢌㒊㡿ᇦ࡛ࡢⓎ⌧ࡢ᭷↓ࢆỴࡵࡿ࠸ ࠺ࡶࡢ࡛࠶ࡿࠋOtx ࡣࠊᕥྑ┦⛠ື≀ࡢ㢌㒊ඹ㏻ ࡋ࡚Ⓨ⌧ࡋ࡚࠸ࡿࡢ࡛ࠊࡇࢀࡽࡢື≀࡛ࡣ Otx ࢆ㢌 㒊㡿ᇦࡢᣦᶆࡋ࡚⏝࠸ࠊࡑࢀࢆᇶಶࠎࡢ㑇ఏᏊ ࡣ CRM ࡢከᵝ㸦type Iࠊtype II ࡞㸧ࢆ㏻ࡌ࡚ 㢌㒊࠾ࡅࡿⓎ⌧ࡢ᭷↓࠾ࡼࡧⓎ⌧㔞ࢆ㑅ᢥࡋࠊࡑ ࡢ⤖ᯝᵝࠎ࡞㢌㒊ࢆ㐍ࡉࡏ࡚ࡁࡓ᥎ᐃࡉࢀࡿࠋ ࡇࢀࡣࠊඹ㏻ࡢไᚚᅉᏊࢆ࠸࡞ࡀࡽࠊ㐍ࡢ㐣⛬ ࡛ᙧែࢆࡁࡃኚ࠼ࡿࡇࡀྍ⬟࡛࠶ࡿࡇࢆ࠺ࡲ ࡃㄝ࡛᫂ࡁࡿࡶࡢ࡛࠶ࡿࠋ 㸰㸧Mix1ࠊVegTࠊSia ࡢ ChIP-seq ゎᯒ ⬇ⴥ≉␗ⓗ⤌⧊ࡸჾᐁࡣศࡋ࡚࠸ࡃࡀࠊࡑࡢ ࡁࢃࢀࡿ enhancer ࡣෆ⬇ⴥᙧᡂࡢࡁணࡵ developmentally poised enhancer ࡋ࡚࢚ࣆࢪ࢙ ࢿࢸࢵࢡ࣐࣮ࢡࡉࢀ࡚࠸ࡿࠊ࠸࠺ࣔࢹࣝࡀᥦ ၐࡉࢀ࡚࠸ࡿࠋࡋࡋࡑࢀࢆᐇドࡋࡓࡣ㠀ᖖᑡ ࡞ࡃࠊࡋࡓࡀࡗ࡚ࡑࡢ୍⯡ᛶࡣ࡛᫂࠶ࡿࠋ㌿ᅉ Ꮚࡢ Mix1 VegT ࡣึᮇࡢෆ⬇ⴥᙧᡂ㛵ࢃࡿࡇ ࡀ♧၀ࡉࢀ࡚࠸ࡿࡀࠊᡃࠎࡣࠊෆ⬇ⴥ⣔ิࡢ developmentally poised enhancer ࡢᙧᡂࡶ㛵ࢃ ࡿࡢ࡛ࡣ࡞࠸ࡢసᴗ௬ㄝࢆࡓ࡚ࠊࡑࡢ᳨ドࢆ⾜ ࡗࡓࠋࡑࡢ⤖ᯝࠊཎ⭠⬇ᮇࡣⓎ⌧ࡋ࡚࠸࡞࠸ࡀࠊ ⚄⤒⬇ᮇࡽᑿⱆ⬇ᮇ௨㝆ࡢෆ⬇ⴥ⣔ิࡢჾᐁ࡛Ⓨ ⌧ࡍࡿ㑇ఏᏊ㏆ഐࡢ CRM Mix1 VegT ࡀ⤖ྜࡋ ࡚࠸ࡿࢆከᩘぢฟࡋࡓࠋ⌧ᅾࡇࢀࡽࡢ CRM ࡀ developmentally poised enhancer ࠺ࢆ࣏ࣞ ࣮ࢱ࣮ゎᯒࡼࡾ᳨ウࡋ࡚࠸ࡿࠋ Sia ࡣࣇࣜ࢝ࢶ࣓࢚࢞ࣝ⬇ࡢ⫼ഃᅉᏊࡢ Wnt ࢩ ࢢࢼ࡛ࣝⓎ⌧ㄏᑟࡉࢀࠊ⫼ഃ࠾ࡼࡧ㢌㒊Ỵᐃࢆ⾜ ࠺㌿ᅉᏊ࡛࠶ࡿࠋࡑࡇ࡛ Sia ࡢᶆⓗ㑇ఏᏊࢆ⥙⨶ ⓗゎᯒࡍࡿࡇ࡛ࠊ㢌㒊ᙧᡂ┤᥋㛵ࢃࡿ㑇ఏᏊ ⩌ࢆྠᐃࡋࠊࡘ Sia ༠ㄪⓗᶵ⬟ࡍࡿ㌿ᅉᏊ ࡢྠᐃࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊSia ࡣ Otx2ࠊMix1ࠊTLEࠊ p300 ࡢࣆ࣮ࢡ㔜࡞ࡿ CRM ࡀ᭱ࡶከࡃ࠶ࡾ㸦type A ྡ㸧ࠊࡘࡇࡢ CRM ࡣ㢌㒊࣮࢜࢞ࢼࢨ࣮㑇 ఏᏊࡢ㏆ഐ᭷ព⃰⦰ࡋ࡚࠸ࡿࡇࡀࢃࡗࡓࠋ ࡉࡽࡡ࣏࣮ࣞࢱ࣮ゎᯒ࡛ࠊtype A CRM ࡣ㢌㒊࣮࢜࢞ ࢼࢨ࣮࡛ࡢⓎ⌧ࢆࡶࡓࡽࡍࡇࡀ♧ࡉࢀࡓࠋ௨ୖ ࡼࡾࠊSia ࡣ type A CRM ࢆࡋ࡚ Otx2ࠊMix1 ༠ㄪⓗ㢌㒊ᙧᡂ㛵ࢃࡿࡇࡀ♧၀ࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ChIP-seq ࡸ RNA-seq ࢆ⮬ࡽ⾜࠺ࡇࡣྍ⬟ࡔࡗ ࡓࡀࠊࠕࢤࣀ࣒ᨭࠖࡼࡾࠊX. tropicalis ⬇ࡑࢀ ࡀ㐺⏝࡛ࡁࡿࢆ᳨ウࡋࠊᐇ㝿ࡇࢀࡽࡢ᪉ἲࡀ㠀ᖖ ᭷ຠ࡛࠶ࡿࡇࡀุ᫂ࡋࠊࡁࡃᒎᮃࢆᣅࡃࡇ ࡀ࡛ࡁࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ㄽᩥⓎ⾲ Yuuri Yasuoka, Yutaka Suzuki, Shuji Takahashi, Haruka Someya, Norihiro Sudou, Yoshikazu Haramoto, Ken Cho, Makoto Asashima, Sumio Sugano, Masanori Taira (2014). Occupancy of Tissue-Specific cis-Regulatory Modules by Otx2 and TLE/Groucho for Embryonic Head Specification. Nat. Commun. 5, 4322 (article number). ᏛⓎ⾲ Yuuri Yasuoka, Yutaka Suzuki, Shuji Takahashi, Haruka Someya, Norihiro Sudou, Yoshikazu Haramoto, Ken Cho, Makoto Asashima, Sumio Sugano, Masanori Taira "Genomics Study of the Spemann-Mangold Organizer: Occupancy of Tissue-Specific cis-Regulatory Modules by Otx2 and TLE/Groucho for Embryonic Head Specification." 㸦Ᏻᒸࠊཱྀ㢌Ⓨ⾲㸹PD ཱྀ㢌Ⓨ⾲㈹ࢆ ཷ㈹㸧 15th International Xenopus Conferenceࠊ Asilomar (California, USA)㸦2014 ᖺ 8 ᭶ 24 ᪥㹼30 ᪥㸧 Yuki Honda, Yoshiaki Kirigaya, Yuuri Yasuoka, Saya Imai, Yutaka Suzuki, Shuji Takahashi, Makoto Asashima, Sumio Sugano, Masanori Taira ͆ ChIP-Seq analysis of Transcription factor Siamois for mechanisms of head organizer formation͇ ➨ 36 ᅇศᏊ⏕≀Ꮫᖺࠊ⚄ᡞ㸦2013 ᖺ 12 ᭶ 3 ᪥㹼6 ᪥㸧 Yuuri Yasuoka, Yutaka Suzuki, Shuji Takahashi, Norihiro Sudou, Yoshikazu Haramoto, Yukiko Tando, Kaoru Kubokawa, Ken W. Cho, Makoto Asashima, Sumio Sugano and Masanori Taira "Otx2 and TLE/Groucho occupancy marks tissue-specific cis-regulatory modules for head specification" ➨ 35 ᅇศᏊ⏕≀Ꮫᖺࠊ ⚟ᒸ㸦2012 ᖺ 12 ᭶ 11 ᪥㹼14 ᪥㸧 Yuuri Yasuoka, Yutaka Suzuki, Shuji Takahashi, Norihiro Sudou, Yoshikazu Haramoto, Yukiko Tando, Kaoru Kubokawa, Ken W. Cho, Makoto Asashima, Sumio Sugano and Masanori Taira "Massively parallel regulation of head and non-head genes by Otx2, Lim1 and Gsc is based on evolution of the head organizer in the chordate" ᪥ ᮏⓎ⏕⏕≀Ꮫ➨ 45 ᅇࠊ⚄ᡞ㸦2012 ᖺ 5 ᭶ 28 ᪥㹼31 ᪥㸧 - 401 - බເᨭㄢ㢟㸸2013 ᖺᗘ RNA ⦅㞟ᵝᘧࡢள✀㛫ᕪࢆ⏝ࡋࡓ࢚ࢿࣝࢠ࣮௦ㅰ㛵㐃࣭㔞ⓗᙧ㉁㑇ఏᏊࡢ᥈⣴ ᇶ┙◊✲㸦㹁㸧 㧗⏣ ㇏⾜࣭ሗ࣭ࢩࢫࢸ࣒◊✲ᶵᵓᅜ❧㑇ఏᏛ◊✲ᡤ ⣔⤫⏕≀◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 NGS ゎᯒࡣሷᇶ㓄ิࡢィ ࢆ㏻ࡋ࡚ࠊࡇࢀࡲ࡛ ྍ⬟࡛࠶ࡗࡓໟᣓⓗ࡞ᝈ㛵㐃㑇ఏᏊࡢ᥈⣴ࢆྍ⬟ ࡋࡘࡘ࠶ࡿࠋࡋ࡚ࠊᮏ◊✲ࡀ┠ⓗࡍࡿ RNA ⦅㞟㛵ࡍࡿ◊✲࡛ࡣࠊࣄࢺࡢ B ⣽⬊ࡢゎᯒࡼࡾࠊ ㌿ᚋࡢ RNA ⦅㞟ࡼࡿ࣑ࣀ㓟⨨ࡀ」ᩘ᳨ฟ ࡉࢀࡓ㸦Li et al. Science 2011㸧ࠋࡲࡓࠊ୰ᯡ⚄⤒⣔ ࡛᭱ࡶάⓎ RNA ⦅㞟ᶵ⬟ࡍࡿ㓝⣲ Ader㸦ࢹ ࣀࢩࣥࢆࣀࢩࣥኚࡍࡿ㸧ࢆᑐ㇟ࡋ࡚ࠊ࣐࢘ ࢫࡢ 15 ⣔⤫ࡢ RNA ⦅㞟ࢆཷࡅࡿሷᇶࡢྠᐃኚ ⋡ࡢẚ㍑ࢆ⾜࠸ࠊࡇࡢ㓝⣲ࡀ㛵ࡍࡿ RNA ⦅㞟ࡢ⣔ ⤫㛫ከᵝᛶࡘ࠸࡚ሗ࿌ࡋࡓ㸦Danecek et al. Genome Biology, 2012㸧࡞ࡶ࠶ࡿࠋᮏ◊✲ࡀᑐ㇟ ࡍࡿ࢚ࢿࣝࢠ࣮௦ㅰ㛵㐃⾲⌧ᆺ RNA ⦅㞟㓝⣲ ࡢ㛵ࢃࡾࡘ࠸࡚ࡣࠊ⾑୰ࡢ⬡㉁㐠ᦙ㔜せ࡞࣏ ࣏ࣜ⺮ⓑ㉁ࡢ㸯ࡘ࡛࠶ࡿ ApoB ࡢ mRNA ࡀ Apobec1 ࡼࡾ⦅㞟ࢆཷࡅࡿࡀ့ங㢮࡛ึࡵ࡚Ⓨぢࡉࢀ࡚ ࠸ࡿ㸦Chen et al. Science, 1987; Powell et al. Cell, 1987㸧ࠋࡲࡓࠊApobec1 ࡢ㑇ఏᏊ◚ቯಶయࡣࠊ⾑୰ࡢ పẚ㔜࣏ࣜ⺮ⓑࢥࣞࢫࢸ࣮ࣟࣝࡢ್ࡀ㢧ⴭୖ᪼ࡍ ࡿࡀ㸦Hirano et al. JBC, 1996; Morrison et al. PNAS, 1996㸧ࠊࡇࢀࡣ Apobec1 ࡢ ApoB ࢆ㐺ṇ࡞ࢧ ࢬࡢ࣏ࣜ⺮ⓑ⩻ヂࡍࡿࡓࡵᚲせ࡞ RNA ⦅㞟 ᶵ⬟ࡀ႙ኻࡍࡿࡓࡵ࡛࠶ࡿࠋ ⏦ㄳ⪅ࡽࡣࠊ ᐇ㦂⏝㏆⣔࣐࢘ࢫ࡛࠶ࡿ C57BL/6J (B6; Mus musculus domesticus)⣔⤫᪥ᮏ⏘㔝⏕ ⏤ ᮶ ࡢ MSM/Ms (MSM; Mus musculus molossinus)⣔⤫ࢆࢶ࣮ࣝࡋ࡚ࠊࣄࢺࡢࠕ㔞ⓗᙧ㉁ࠖ 㛵ࢃࡿ㑇ఏᅉᏊࡸࡑࢀࡽࡀ㛵ࢃࡿࢿࢵࢺ࣮࣡ࢡࢆ ༢㞳࣭ྠᐃࡍࡿࡓࡵࡢ◊✲ࢆ⾜ࡗ࡚࠸ࡿࠋ≉ࠊB6 ࡢ㑇ఏⓗ⫼ᬒ MSM ࡢᰁⰍయࢆࡈ୍ᮏࡎࡘ⨨ ࡋ࡚ᶞ❧ࡋࡓ࣐࢘ࢫள✀㛫ࢥࣥࢯ࣑ࢵࢡ㸦ᰁⰍయ ⨨㸧⣔⤫ࡢつᶍ⾲⌧ᆺゎᯒࠊ⤖ᯝࡢࢹ࣮ࢱ࣋ ࣮ࢫࢆ⾜࠸ࠊ୍㒊ࢆබ㛤ࡋ࡚࠸ࡿ㸦Takada et al. Genome Res. 2008; Takada & Shiroishi, Exp. Anim. 2012㸧ࠋᮏ⏦ㄳㄢ㢟ࡀᑐ㇟ࡍࡿ࢚ࢿࣝࢠ࣮௦ ㅰ㛵㐃⾲⌧ᆺࡘ࠸࡚ࡣࠊྛ✀ࡢࢥࣥࢯ࣑ࢵࢡ⣔⤫ ࢆ B6 ẚ㍑ࡋ࡚ࠊయ㔜ࠊ⬡⫫⋡㸦య㔜ᑐࡍࡿయ⬡ ⫫ྜ㸧 ࠊ࠾ࡼࡧ⾑୰ࡢྛ✀ࢥࣞࢫࢸ್࣮ࣟࣝࢆኚື ࡉࡏࡿ㑇ఏᅉᏊࡀᏑᅾࡍࡿᰁⰍయࢆ」ᩘ᳨ฟࡋ࡚࠸ ࡿ(Takada et al. Genome Res. 2008)ࠋࡉࡽࠊ⏦ㄳ ⪅ࡽࡣ MSM ⣔⤫ࡢࢤࣀ࣒ࢆࣜࢩ࣮ࢡ࢚ࣥࢫࡋ࡚ࠊ ࢤࣀ࣒ከᆺሗࢆබ㛤ࡋ࡚࠸ࡿࡀ㸦Takada et al. Genome Res. 2013; Takada et al. Mammal. Genome 2015㸧ࠊB6-MSM 㛫ࡣࠊᮏ⏦ㄳㄢ㢟ࡀᑐ ㇟ࡋ࡚࠸ࡿ RNA ⦅㞟㛵ࡍࡿ」ᩘࡢ㓝⣲ ࣑ࣀ㓟⨨ࡀᏑᅾࡍࡿࡇࡀほᐹࡉࢀ࡚࠸ࡿࠋᮏ⏦ ㄳ࡛ࡣࡇࡢࡼ࠺࡞≧ἣࢆࡩࡲ࠼ࠊ࣐࢘ࢫࡢ࢚ࢿࣝࢠ ࣮௦ㅰ㛵ಀࡋࡓ㔞ⓗᙧ㉁ࡢศᏊᇶ┙ RNA ⦅㞟 ࡀ㛵ࢃࡗ࡚࠸ࡿࡢ࡛ࡣ࡞࠸࠸࠺௬ㄝࢆᚓࠊࡇࢀ ࢆ᳨ドࡍࡿ࡛ࠊ࢚ࢿࣝࢠ࣮௦ㅰ⾲⌧ᆺ㛵ࢃࡿ RNA ⦅㞟㓝⣲ࡢከᵝᛶࡢ༢㞳࣭ྠᐃ⧅ࡆࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᮏᨭㄢ㢟࡛ࡣࠊ࣐࢘ࢫࡢࢥࣥࢯ࣑ࢵࢡ⣔⤫⩌ ࡽࠊయ㔜ࡸ⾑୰ࡢྛ✀ࢥࣞࢫࢸ್࣮ࣟࣝ࡞ࠊ࢚ࢿ ࣝࢠ࣮௦ㅰ㛵㐃⾲⌧ᆺ≉ᚩࡢ࠶ࡿ⣔⤫ࠊ࠾ࡼࡧࢥ ࣥࢺ࣮ࣟࣝࡋ࡚ B6 ࠾ࡼࡧ MSM ⣔⤫ࡢ⫢⮚ࢧࣥࣉ ࣝࢆᑐ㇟ࠊNGS ࡼࡿ㑇ఏᏊⓎ⌧ゎᯒࡢᨭࢆཷ ࡅࡓࠋලయⓗ࡞ᨭෆᐜࡣࠊ࣐࢘ࢫᐇ㦂⣔⤫ B6ࠊ MSM ࠾ࡼࡧࢥࣥࢯ࣑ࢵࢡ⣔⤫ 2 ✀㸦4 ⣔⤫㸧ࡢ㞤㞝 ྛ 3 㢌ࠊィ 24 ✀ࡢ⫢⮚ࡼࡾ⢭〇ࡋࡓ RNA ヨᩱࢆ ࣑ࣝࢼ♫ HiSeq ࡾゎᯒࡋࡓࢹ࣮ࢱ࡛࠶ࡿ㸦࢚࣌ ࣥࢻ 100bpࠊ30-70M ࣮ࣜࢻࠊQV30: >93.26%)ࠋࡲ ࡓࠊࡇࢀࡽࡢࢹ࣮ࢱࡢ DDBJ㸦DRA㸧ࡢⓏ㘓ࡘ ࠸࡚ࡶࡈᨭ࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏᨭࡼࡾཷࡅࡓ⤖ᯝࡢ 2 ḟゎᯒࡽࠊ㑇ఏᏊ Ⓨ⌧ࡢ⣔⤫㛫ᕪ࠾ࡼࡧ RNA ⦅㞟ཷᐜೃ⿵ SNP 㛵 ࡍࡿሗࢆᚓࡓࠋࡇࡢ⤖ᯝࡣࠊ᪤▱ࡢ RNA ⦅㞟ཷ ᐜሷᇶࢆ᳨ฟࡍࡿࡇࡀ࡛ࡁ࡚࠸ࡿࠋB6 ࠾ࡼࡧ MSM ࡶࠊⓎ⌧㑇ఏᏊࡢ RNA ⦅㞟ཷᐜೃ⿵㑇ఏᏊ ᩘࡣ㸪Ⓨ⌧ኚື㑇ఏᏊ㸦DEG㸧ࡀⓎ⌧ኚືࡢ↓࠸㑇 ఏᏊ㸦non-DEG㸧ࡼࡾࡶ 2 ಸ௨ୖከࡗࡓࠋࡉࡽࠊ 㑇ఏᏊᶵ⬟㸦GO Term㸧㛵ࡍࡿሗゎᯒࢆ⾜ࡗࡓ ⤖ᯝࠊ㌿ࡸ௦ㅰࠊࡉࡽࡣჾᐁⓎ⏕ࡢㄪ⠇㛵ࡍ ࡿ㑇ఏᏊ⩌ࡀほᐹࡉࢀࡓࠋ⌧ᅾࡇࢀࡽࡀ B6 ࠾ࡼࡧ MSM ࡢ࢚ࢿࣝࢠ࣮௦ㅰ⾲⌧ᆺࡢ⣔⤫㛫ᕪ㛵ࢃࡿ ࢆྛ✀ᐇ㦂ࡼࡾ᳨ウࡋ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣐࢘ࢫಶయ࣮࣋ࢫࡢ㑇ఏᏊⓎ⌧ゎᯒࡘ࠸࡚ࡣࠊ ࡇࢀࡲ࡛ࠕ㑇ఏᏊⓎ⌧㔞ࡢ㐪࠸ࠖࠕࢤࣀ࣒㓄ิୖ ࡢⓎ⌧㒊ࠖࡢ᥈⣴◊✲ࡀ࡛࠶ࡗࡓࡀࠊ ࠕࢤࣀ࣒ᨭ ࠖࢆά⏝ࡋࡓ㧗⢭⣽࡞ゎᯒࡼࡾࠊ ࠕ⩻ヂࡉࢀࡿࢱ ࣥࣃࢡ㉁ኚࢆࡶࡓࡽࡍ RNA ⦅㞟ࡢள✀㛫ᕪࠖ ࡘ࠸࡚ࡶ⥙⨶ⓗ᥈⣴◊✲ࢆ⾜࠺ࡇࡀྍ⬟࡞ࡗ ࡓࠋࡲࡓࠊᮏ◊✲࡛⾜ࡗࡓ㑇ఏᏊⓎ⌧ゎᯒࡽࠊ࣐ ࢘ࢫࡢ࢚ࢿࣝࢠ࣮௦ㅰ⾲⌧ᆺࡢᕪ␗ࠊࢤࣀ࣒⦅㞟 ࡢள✀㛫ᕪࡀ㛵ࢃࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 402 - ࡞ࡋ බເᨭㄢ㢟㸸2014 ᖺᗘ ࣔࢹ࣐ࣝ࢘ࢫࢆ⏝࠸ࡓ᪂つᦤ㣗࣭⬡⫫✚ㄪ⠇㑇ఏᏊࡢᶵ⬟ゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ 㧗⏣ ㇏⾜࣭ሗ࣭ࢩࢫࢸ࣒◊✲ᶵᵓᅜ❧㑇ఏᏛ◊✲ᡤ ⣔⤫⏕≀◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ࣐࢘ࢫ㸦Mus musculus㸧ࡣࠊࣄࢺࡢࡉࡲࡊࡲ࡞ య㉁ࡸᝈ㢮ఝࡋࡓ⾲⌧ᆺࢆ♧ࡍከᩘࡢᐇ㦂⏝⣔ ⤫ࡀᏑᅾࡋࠊᝈ㛵㐃ᅉᏊࡢ᥈⣴ጾຊࢆⓎࡋ࡚ ࠸ࡿࠋ⏦ㄳ⪅ࡽࡣࠊࡇࡢᐇ㦂ື≀ࠕ࣐࢘ࢫࠖࢆࢶ࣮ ࣝࡋ࡚ࠊయ㔜ࡸ࠶ࡿ✀ࡢᝈᑐࡍࡿឤཷᛶ࡞ ࠕ㔞ⓗᙧ㉁ࠖࡢཎᅉ㑇ఏᏊࡸࠊࡑࢀࡽࡀᶵ⬟ࡍࡿࢿ ࢵࢺ࣮࣡ࢡࢆ༢㞳࣭ྠᐃࡍࡿࡓࡵࡢ◊✲ࢆ⾜ࡗ࡚࠸ ࡿࠋࡇࢀࡲ࡛ࠊᐇ㦂⏝㏆⣔࣐࢘ࢫ࡛࠶ࡿす࣮ࣚ ࣟ ࢵ ࣃ ⏘ ள ✀ ࡢ C57BL/6J (B6; Mus musculus domesticus) ⣔ ⤫ ᪥ ᮏ ⏘ ள ✀ ࡢ MSM/Ms(MSM; Mus musculus molossinus)⣔⤫ࢆ⏝ࡋ࡚ᶞ❧ࡉ ࢀࡓ࣐࢘ࢫள✀㛫ࢥࣥࢯ࣑ࢵࢡ㸦ᰁⰍయ⨨㸧⣔⤫ ⩌ࢆ⏝ࡋ࡚ࠊ࢚ࢿࣝࢠ࣮௦ㅰ㛵㐃ᙧ㉁ࡢࢫࢡ࣮ࣜ ࢽࣥࢢࡼࡿໟᣓⓗ࡞⾲⌧ᆺゎᯒࢆ⾜ࡗࡓ⤖ᯝࠊ㐣 㣗࣭⫧‶ࢆ㉳ࡇࡍࢥࣥࢯ࣑ࢵࢡ⣔⤫ࢆⓎぢࡋࡓ 㸦Takada et al. Genome Res. 2008㸧ࠋ᭱㏆ࠊࡇࡢ㐣 㣗࣭⫧‶ࢆ㉳ࡇࡍཎᅉࡀࠊ㸯✀㢮ࡢ㑇ఏᏊ㉳ࡁࡓ ሷᇶ⨨㉳ᅉࡍࡿྍ⬟ᛶࡀ㧗࠸ࡇࢆ✺ࡁṆࡵࡓ 㸦ᢞ✏‽ഛ୰㸧 ࠋࡇࡢ㑇ఏᏊࡣ࣐࢘ࢫࡢࡳ࡞ࡽࡎࠊࣄ ࢺࢆࡣࡌࡵࡋࡓ့ஙື≀ᗈࡃಖᏑࡉࢀ࡚࠸ࡿࡀࠊ ࢹ࣮ࢱ࣮࣋ࢫୖࡢྡ⛠ࡀ࠼ࡽࢀ࡚࠸ࡿࡶࡢࡢࠊᶵ ⬟㛵ࡍࡿሗ࿌ࡣ↓࠸ࠋࡑࡇ࡛ࠊᙜヱ㑇ఏᏊࡢᶵ⬟ ࢆ᫂ࡽࡍࡿࡶࠊሷᇶ⨨ࡼࡿᶵ⬟ኚ ࡀࠊ㐣㣗࣭⫧‶ࢆᘬࡁ㉳ࡇࡍཎᅉࢆゎ᫂ࡍࡿࡇ ࡼࡾࠊࣄࢺࡢᦤ㣗␗ᖖࡸࡇࢀࡀཎᅉ࡛㉳ࡇࡿ⫧‶ࠊ ࡉࡽࡣ㛵㐃ࡍࡿᝈ◊✲ᙺ❧ࡘሗࢆᚓࡿࡇ ࡀ࡛ࡁࡿ⪃࠼ࡓࠋ⏦ㄳ⪅ࡢࡇࢀࡲ࡛ࡢ◊✲ࡽࠊ ᙜヱ㑇ఏᏊࢆᑐ㇟ࡋࡓ࣏࣮ࣞࢱ࣮㑇ఏᏊࡢⓎ⌧ࢆ ほᐹࡋࡓࡇࢁࠊ⬻ࡢ㛫⬻ࡽᘏ㧊ࡅ࡚ࡢᗈ࠸ ⠊ᅖ࡛㑇ఏᏊࡢⓎ⌧ࡀண ࡉࢀࡓࠋḟࠊබඹࡢࢹ ࣮ࢱ࣮࣋ࢫࢆ⏝ࡋ࡚ࠊᙜヱ㑇ఏᏊࡢⓎ⌧ືែࢆ☜ ㄆࡋࡓࡇࢁࠊᩘ✀㢮㸦ᩘⓒࠥ2 ༓ bp㸧ࡢ㌿⏘≀ ࡀሗ࿌ࡉࢀ࡚࠸ࡓࠋ⏦ㄳ⪅ࡢࡇࢀࡲ࡛ࡢゎᯒࡽࠊ ⬻࠾ࡅࡿᶆグ㑇ఏᏊࡢⓎ⌧ࡣࠊB6 ẚ㍑ࡋ࡚ࠊࢥ ࣥࢯ࣑ࢵࢡ⣔⤫࠾ࡼࡧ㑇ఏᏊ◚ቯ࣐࢘ࢫࡶప࠸ ࡇࡀ☜ㄆ࡛ࡁ࡚࠸ࡿࡀࠊMSM ⣔⤫⏤᮶ࣞࣝࢆᣢ ࡘᶆグࢥࣥࢯ࣑ࢵࢡ⣔⤫࠾ࡅࡿࡑࡢࣂ࢚࣮ࣜࢩࣙ ࣥࡣ࡛᫂࠶ࡿࠋᮏᨭ࡛ࡣࠊᙜヱ㑇ఏᏊࡢᶵ⬟ࡸࠊ ࡑࡢⓎ⌧㛵ࡍࡿゎᯒࢆ⾜࠺ࡓࡵࠊPacBioRS ll ௦ ⾲ࡉࢀࡿࠊ୍ศᏊࣜࣝࢱ࣒ DNA ࢩ࣮ࢣࣥࢧࡼ ࡿ㛗 cDNA ࡢゎㄞࢆᕼᮃࡋࡓࠋࡇࢀࡼࡾࠊᦤ 㣗͌⫧‶㛵ࢃࡿᙜヱ㑇ఏᏊࡢࢫࣉࣛࢩࣥࢢࣂࣜ ࢚࣮ࢩࣙࣥࡢ᳨ฟࠊṇᖖኚ␗ಖᣢ࣐࢘ࢫ࡛⬻࠾ ࠸࡚Ⓨ⌧ࡀኚືࡋ࡚࠸ࡿ㑇ఏᏊࠊࡉࡽࡣࡑࢀࡽࡀ ᒓࡍࡿࣃࢫ࢙࢘࡞ࠊ㐣㣗͌⫧‶㛵ࢃࡿ⬻ෆ࡛ࡢ 㛵㐃㑇ఏᏊⓎ⌧ࢿࢵࢺ࣮࣡ࢡࡢྠᐃࡀᮇᚅ࡛ࡁࠊᙜ ヱ㑇ఏᏊࡢᶵ⬟ࡑࡢ◚⥢ࡼࡿ⫧‶Ⓨࡢཎᅉࢆ ᫂ࡽ࡛ࡁࡿ⪃࠼ࡓࠋ cDNA ゎㄞࢆ MSMࠊB6ࠊࢥࣥࢯ࣑ࢵࢡ⣔⤫ࠊ㑇ఏ Ꮚ◚ቯ⣔⤫ࡢ㞤㞝ࠊィ 8 ✀ࡘ࠸࡚⾜ࡗ࡚࠸ࡓࡔ࠸ ࡓࠋࢧࣥࣉࣝࡣ 10 㐌௧ࡢື≀ࢆᑐ㇟ࡋࠊ⬻ࡼࡾ⢭ 〇ࡋࡓ RNA ࡽ㛗 cDNA ࢆస〇ࡋゎᯒ౪ࡋ ࡓࠋᮏᨭ࡛ᑐ㇟ࡍࡿ㑇ఏᏊࡢ㌿⏘≀ࡣࠊ࠾࠾ ࡴࡡ 2-3kb ண ࡉࢀࡓࡢ࡛ࠊྛࢧࣥࣉࣝᑐࡋ࡚ࠊ 1-3kb ࠾ࡼࡧ 3-6kb ࡢ㛗 cDNA ࢆᑐ㇟ࡋࡓ 2 ✀㢮ࡢࣛࣈ࣮ࣛࣜࢆస〇࠸ࡓࡔࡁࠊࡑࢀࡒࢀࢆゎ ᯒࡋ࡚࠸ࡓࡔ࠸ࡓࠋᚓࡽࢀࡓゎᯒ⤖ᯝࡣࠊࣇࣝࢱ ࣜࣥࢢ๓ࡢ≧ែ࡛ࠊࢧࣥࣉࣝ㸯✀ࡘࡁࠊ୧᪉ࡢࣛ ࣈ࣮ࣛࣜࢆྜࢃࡏ࡚ᖹᆒ 60 ࣮ࣜࢻࡘ࠸࡚ࡢ ゎᯒሗࢆ࠸ࡓࡔ࠸ࡓࠋࡲࡓࠊࡇࢀࡽࡢࢹ࣮ࢱࡽ ᙜヱ㑇ఏᏊࢆྵࡴⓎ⌧㑇ఏᏊ᳨ฟࡢࡓࡵࡢ࣐ࢵࣆࣥ ࢢࡸࠊ㛗 cDNA ࡢ㓄ิᩘࢆྵࡴሗゎᯒࡘ࠸࡚ ࡶ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᮏ⏦ㄳ࡛ࡣࠊ PacBio ࢩ࣮ࢣࣥࢧࡼࡿ㛗 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏᨭࡼࡾゎᯒ࠸ࡓࡔ࠸ࡓ⤖ᯝࡽࠊᙜヱ㑇ఏ Ꮚ㛵ࡋ࡚ࠊࡇࢀࡲ᳨࡛ドࡀ㠀ᖖᅔ㞴ࡔࡗࡓ MSM ⣔⤫ࡢࢫࣉࣛࢩࣥࢢࣇ࢛࣮࣒ࡢࣂ࢚࣮ࣜࢩࣙࣥࠊ ྛ✀ື≀࠾ࡅࡿ」ᩘ㑇ఏᏊࡢ㑇ఏᏊⓎ⌧ືែࡢᴫ せࠊṇᖖኚ␗ಖᣢࠊࡉࡽࡣ㑇ఏᏊ◚ቯື≀㛫ࡢ ⬻࡛Ⓨ⌧ࡀኚືࡍࡿ㑇ఏᏊ⩌ࡢ୍㒊ࡀ᫂ࡽ࡞ࡗ ࡓࠋ⌧ᅾࠊྛ⣔⤫ࡢࢫࣉࣛࢩࣥࢢࣇ࢛࣮࣒ࡢ᳨ドࠊ Ⓨ⌧ࡀኚືࡋ࡚࠸ࡿ㑇ఏᏊࡀ㛵ࢃࡿࣃࢫ࢙࢘ࡢ᥈ ⣴࡞ࠊ㐣㣗͌⫧‶㛵ࢃࡿ⬻ෆ࡛ࡢ㛵㐃㑇ఏᏊⓎ⌧ ࢿࢵࢺ࣮࣡ࢡࡢྠᐃࢆ㐍ࡵ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡾࠊ⫧‶ࡢཎᅉ㑇ఏᏊ⪃࠼ࡽࢀࡿᙜ ヱ㑇ఏᏊࡘ࠸࡚ࠊࡇࢀࡲ࡛ࡣ㠀ᖖᅔ㞴࡛࠶ࡗࡓ 」ᩘࡢࢫࣉࣛࢩࣥࢢࣇ࢛࣮࣒ࡢྠᐃࢆ୍ศᏊࢩ࣮ ࢣࣥࢩࣥࢢࡼࡾ⾜࠺ࡇࡀ࡛ࡁࡓࠋࡋࡶࠊ㛵㐃 ࡍࡿゎᯒࣀ࢘ࣁ࢘ࢆᵓ⠏࡛ࡁࡓࡇࡶࡁ࡞ᡂᯝ࡛ ࠶ࡿࠋࡋࡋ࡞ࡀࡽࠊᑐ㇟㑇ఏᏊࡢ㌿⏘≀ࡀᑡ࡞ ࠸ࡇࡶ࠶ࡾࠊㄽᩥࡢ㝿ࡣࡉࡽከࡃࡢ㌿⏘ ≀ࢆゎᯒࡍࡿᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡿࡇࡽࠊᶆ ⓗ㑇ఏᏊࡢ cDNA ࣛࣈ࣮ࣛࣜࡽࡢ⃰⦰࡞ࡼ ࡿゎᯒࡢᚲせࡀ࠶ࡿᛮࢃࢀࡿࠋMSM ⣔⤫ࡢᙜヱ㑇 ఏᏊࡘ࠸࡚ࡶ」ᩘࡢ㑅ᢥⓗࢫࣉࣛࢩࣥࢢࣇ࢛࣮ ࣒ࡀᏑᅾࡋ࡚࠸ࡓࡓࡵࠊࢱࣥࣃࢡ㉁ࡣྜᡂࡉࢀ࡞࠸ ࡶࡢࡢࠊRNA ࡣ㌿ࡉࢀ࡚࠸ࡿࡇࡀ☜ㄆ࡛ࡁࡓࠋ ࡉࡽࠊ㑇ఏᏊ◚ቯື≀ࡣࠊᙜヱ㑇ఏᏊ㛵ࡍࡿ㌿ ࢆ☜ㄆ࡛ࡁ࡞ࡗࡓࡀࠊࡇࢀࡣゎᯒ࣮ࣜࢻᩘࡀ ẚ㍑ࡋ࡚༙ศ⛬ᗘ࡞ࡢࡀᙳ㡪ࡋ࡚࠸ࡿྍ⬟ᛶࡶ⪃ ࠼ࡽࢀࡿࠋᚋࡣࠊᮏᨭࡼࡿ⤖ᯝຍ࠼࡚ࠊ࣐ ࢡࣟࣞࡸ▷㙐୪ิᆺࢩ࣮ࢣࣥࢧ࡞ࡼࡿ㑇 ఏᏊⓎ⌧ゎᯒࢆ⾜࠸ࠊ⿵ⓗ࡞ゎᯒࢆ㐍ࡵࡿᚲせࡀ ࠶ࡿࡇࡶ☜ㄆ࡛ࡁࡓࠋ - 403 - බເᨭㄢ㢟㸸2013 ᖺᗘ ᪂つ㌿ఙ㛗ไᚚᅉᏊ㹋㹃㹂㸰㸴ࡢᶆⓗ㑇ఏᏊゎ᫂ᛴᛶⓑ⾑ࡢ㛵ࡘ࠸࡚ࡢゎᯒ ⱝᡭ◊✲㸦㸿㸧 㧗ᶫ ⚽ᑦ࣭ᾏ㐨ᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ከࡃࡢࣄࢺ㑇ఏᏊ࡛ RNA ࣏࣓࣮ࣜࣛࢮ II㸦Pol II㸧 ࡀ㌿㛤ጞ┤ᚋ୍Ṇࡋ࡚࠸ࡿࡇࡀࢃࡾࠊ 㑇ఏᏊⓎ⌧࠾࠸࡚㌿ఙ㛗ࡢ㐣⛬ࡀ㔜せ࡞ᙺࢆ ᯝࡓࡋ࡚࠸ࡿࡇࡀࢃࡗ࡚ࡁࡓࠋPol II ࡢ୍Ṇ ࡀゎ㝖ࡉࢀࠊPol II ࡀ RNA ྜᡂࢆ㛤ࡍࡿࡓࡵ ࡣࠊ㌿ఙ㛗ᅉᏊࡀ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡉࢀࠊ Pol II ࡢ୍Ṇࢆゎ㝖ࡍࡿᚲせࡀ࠶ࡿࠋ⏦ㄳ⪅ࡣ࣓ ࢹ࢚࣮ࢱ࣮」ྜయࡢࢧࣈࣘࢽࢵࢺ Med26 ࡀ㌿ ఙ㛗ᅉᏊ」ྜయ͆Super elongation complex͇ 㸦SEC㸧 ࢆ c-Myc ࡸ Hsp70 ࡞ࡢ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡋࠊ ࡑࢀࡽࡢ㌿ఙ㛗ࢆಁ㐍ࡍࡿࡇࢆ᫂ࡽࡋࡓ ࠙Takahashi H, et al. Cell 2011ࠚࠋࡉࡽࠊ⏦ㄳ⪅ ࡣ Med26 ⤖ྜࡍࡿࡶ࠺୍ࡘࡢ㌿ఙ㛗ᅉᏊ」ྜ య͆Little elongation complex͇ 㸦LEC㸧ࢆྠᐃࡋࡓࠋ ᮏ◊✲࡛ࡣ Med26 ࡀ SEC LEC ࢆࡑࢀࡒࢀࡢࡼ ࠺࡞㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡍࡿࡢࡘ࠸࡚᫂ࡽ ࡍࡿࠋ ࡲࡓࠊ⯆῝࠸ࡇࠊSEC ࡢࢧࣈࣘࢽࢵࢺࡢ ELLࠊAF4ࠊAFF4ࠊAF9 ࡸ ENL ࡢ㑇ఏᏊࡣ MLL 㸦Mixed Lineage Leukemia㸧㑇ఏᏊΰྜᆺᛴᛶⓑ ⾑࡛㧗㢖ᗘᰁⰍయ㌿ᗙࡀࡳࡽࢀࡿࠋ᭱㏆ࡢ◊✲ ࡛ࠊ㌿ᗙࡢ⤖ᯝ⏕ࡌࡓ MLL ⼥ྜࢱࣥࣃࢡ㉁ࡼࡗ࡚ SEC ࡀ Hox ࡞ࡢⓑ⾑㛵㐃㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝ ࢺࡉࢀࠊࡑࡢ㑇ఏᏊࡢⓎ⌧ࢆ␗ᖖஹ㐍ࡉࡏࡿࡇ ࡀΰྜᆺᛴᛶⓑ⾑ࡢⓎ࣓࢝ࢽࢬ࣒ࡢ 1 ࡘ࡛࠶ࡿ ࡇࡀࢃࡗࡓࠋࡇࡢࡇࡽࠊΰྜᆺᛴᛶⓑ⾑ ࠾࠸࡚ࠊ࣓ࢹ࢚࣮ࢱ࣮」ྜయࡀ Pol II ඹ 㸦Med26 SEC ࡢ⤖ྜࢆࡋ࡚㸧ⓑ⾑㛵㐃㑇 ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡉࢀࠊࡑࢀࡽࡢⓎ⌧ࢆஹ㐍ࡉ ࡏ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋᮏ◊✲࡛ࡣΰྜᆺᛴ ᛶⓑ⾑࠾࠸࡚ Med26 ࡀ Pol II ඹⓑ⾑㛵㐃 㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡉࢀࡿࡢࡘ࠸࡚ࡶ᫂ࡽ ࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ەHEK293T ⣽⬊ࢆ⏝࠸ࡓ Med26ࠊAFF4㸦SEC ࡢ ࢧࣈࣘࢽࢵࢺ㸧 ࠊKIAA0947㸦LEC ࡢࢧࣈࣘࢽࢵࢺ㸧 ࡢ ChIP㸦ࢡ࣐ࣟࢳࣥචỿ㝆㸧ࢩ࣮ࢡ࢚ࣥࢫゎᯒࠋ ە㔝⏕ᆺࡢ MLL ࢆⓎ⌧ࡍࡿ⣽⬊㸦REH ⣽⬊㸧 MLL-SEC ࢧࣈࣘࢽࢵࢺࡢ⼥ྜࢱࣥࣃࢡ㉁ࢆⓎ⌧ࡍ ࡿΰྜᆺᛴᛶⓑ⾑⣽⬊㸦RS4-11 ⣽⬊㸸MLL-AF4 ࢆⓎ⌧㸧 ࢆ⏝࠸ࡓ Med26 ࡢ ChIP ࢩ࣮ࢡ࢚ࣥࢫゎᯒࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㸦1㸧Med26 ࡢ ChIP-seq ゎᯒࡼࡗ࡚ࠊMed26 ࡀ Ꮡᅾࡍࡿ㑇ఏᏊ㡿ᇦࡀ⥙⨶ⓗ᫂ࡽ࡞ࡗࡓࠋࡑ ࢀࡽࡢ㑇ఏᏊࡢ୰࡛ࠊࢱࣥࣃࢡ㉁ࢆࢥ࣮ࢻࡍࡿ㑇ఏ Ꮚࡣ SEC ࡼࡗ࡚ไᚚࡉࢀࠊsmall nuclear RNA 㸦snRNA㸧࡞ࡢ non-coding RNA 㑇ఏᏊࡣ LEC ࡼࡗ࡚ไᚚࡉࢀࡿࡇࡀ♧၀ࡉࢀࡓࠋ 㸦2㸧LEC ࡢࢧࣈࣘࢽࢵࢺࡢ KIAA0947 ࡢ ChIP-seq ゎᯒࡼࡗ࡚ࠊKIAA0947 ࡀᏑᅾࡍࡿ㑇ఏᏊ㡿ᇦࡀ ⥙⨶ⓗ᫂ࡽ࡞ࡗࡓࠋᮏゎᯒࡼࡗ࡚ࠊLEC ࡀ snRNA 㑇ఏᏊࡸࡢ non-coding RNA 㑇ఏᏊ㡿ᇦ Ꮡᅾࡍࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ 㸦3㸧TFIID ࡢࢧࣈࣘࢽࢵࢺࡢ TAF7 ࡢ ChIP-seq ゎ ᯒࡼࡗ࡚ࠊTAF7 ࡀᏑᅾࡍࡿ㑇ఏᏊ㡿ᇦࡀ᫂ࡽ ࡞ ࡗ ࡓ ࠋ ᮏ ゎ ᯒ ࡼ ࡗ ࡚ ࠊ TAF7 ࡣ Med26 ࡸ KIAA0947㸦LEC ࡢࢧࣈࣘࢽࢵࢺ㸧ඹ snRNA 㑇ఏᏊ㡿ᇦᏑᅾࡍࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ HEK293T ⣽⬊ࢆ⏝࠸࡚ TAF7 ࢆࣀࢵࢡࢲ࢘ࣥࡍࡿ ࠊsnRNA 㑇ఏᏊ㡿ᇦ࠾ࡅࡿ LEC ࡢࣜࢡ࣮ࣝࢺ ࡢቑຍࡀࡳࡽࢀࡓࡇࡽࠊTAF7 ࡣ Med26 ࡼࡿ LEC ࡢࣜࢡ࣮ࣝࢺࢆ㜼ᐖࡍࡿࡇࡀ♧၀ࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏゎᯒࡼࡗ࡚ࠊMed26 TAF7 ࡢᶆⓗ㑇ఏᏊࡀ᫂ ࡽ࡞ࡗࡓࠋMed26 TAF7 ඹ㏻ࡍࡿᶆⓗ㑇ఏ ᏊࡣࠊSEC LEC ࡢᶆⓗ㑇ఏᏊࢆඹྵࢇ࡛࠸ࡿ ࡇࡀண ࡉࢀࡓࠋᮏࢤࣀ࣒ᨭࡼࡗ࡚ᚓࡓ⤖ᯝ ࢆᇶࠊMed26 ࡀࡢࡼ࠺ࡋ࡚ࠊSEC LEC ࢆ ␗࡞ࡿ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡍࡿࡢ㛵ࡍࡿ࣓ ࢝ࢽࢬ࣒ࡢゎᯒࢆ⾜࠺ࠋࡉࡽࠊTAF7 ࡀࠊMed26 ࡼࡿ SEC ࡸ LEC ࡢࣜࢡ࣮ࣝࢺࢆ㜼ᐖࡍࡿࡢ 㛵ࡋ࡚ࠊࡉࡽゎᯒࢆ⾜࠸ࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Takahashi H., Takigawa I., Watanabe M., Anwar D., Shibata M., Tomomori-Sato C., Sato S., Ranjan A., Seidel CW., Tsukiyama T., Mizushima W., Hayashi M., Ohkawa Y., Conaway JW., Conaway RC., Hatakeyama S.: MED26 regulates the transcription of snRNA genes through the recruitment of little elongation complex. Nature Commun., 6, 5941, 2015. ەHEK293T ⣽⬊ࢆ⏝࠸ࡓ Med26ࠊAFF4㸦SEC ࡢ ࢧࣈࣘࢽࢵࢺ㸧 ࠊKIAA0947㸦LEC ࡢࢧࣈࣘࢽࢵࢺ㸧ࠊ TAF7 ࡢ ChIP㸦ࢡ࣐ࣟࢳࣥචỿ㝆㸧ࢩ࣮ࢡ࢚ࣥࢫ ゎᯒࠋ - 404 - බເᨭㄢ㢟㸸2014 ᖺᗘ Med26 ࡼࡗ࡚ࣜࢡ࣮ࣝࢺࡉࢀࡿ᪂つ㌿ఙ㛗」ྜయ LEC ࡢᶵ⬟ゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ 㧗ᶫ ⚽ᑦ࣭ᾏ㐨ᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 㠀ᖖከࡃࡢࣄࢺ㑇ఏᏊ࡛ RNA ࣏࣓࣮ࣜࣛࢮ II 㸦Pol II㸧ࡀ㌿㛤ጞ┤ᚋ୍Ṇࡋ࡚࠸ࡿࡇࡀ ࢃࡾࠊ㑇ఏᏊⓎ⌧࠾࠸࡚㌿ఙ㛗ࡢ㐣⛬ࡀ㔜せ ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿࡇࡀࢃࡗࡓࠋPol II ࡀ୍ Ṇࢆゎ㝖ࡉࢀࠊRNA ྜᡂࢆ㛤ࡍࡿࡓࡵࡣࠊ㌿ ఙ㛗ᅉᏊࡀ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡉࢀࡿᚲせ ࡀ࠶ࡿࠋ⏦ㄳ⪅ࡣ࣓ࢹ࢚࣮ࢱ࣮」ྜయࡢࢧࣈࣘࢽ ࢵࢺ Med26 ࡀࠊࡑࡢ N ᮎ➃ࢻ࣓ࣥ㸦NTD㸧ࡼ ࡗ ࡚ ࠊ 2 ࡘ ࡢ ␗ ࡞ ࡿ ㌿ ఙ 㛗 ᅉ Ꮚ 」 ྜ య Super elongation complex 㸦 SEC 㸧 Little elongation complex㸦LEC㸧ࢆࠊࡑࢀࡒࢀ␗࡞ࡿ㑇ఏᏊ㡿ᇦ ࣜࢡ࣮ࣝࢺࡍࡿࡇࢆ᫂ࡽࡋࡓࠋᮏ◊✲࡛ࡣࠊ Med26 ࡀࡢࡼ࠺࡞࣓࢝ࢽࢬ࣒࡛ࠊSEC LEC ࢆ ࡑࢀࡒࢀ␗࡞ࡿ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡍࡿࡢ ࡘ࠸࡚᫂ࡽࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ەMed26 ࡀࡢࡼ࠺࡞㑇ఏᏊࡢไᚚ㛵ࢃࡗ࡚࠸ࡿ ࡢࢆ᫂ࡽࡍࡿࡓࡵࠊControl ࡢ siRNAࠊ Med26 ࡢ siRNA ࢆࡑࢀࡒࢀࢺࣛࣥࢫࣇ࢙ࢡࢩࣙࣥ ࡋࡓ HEK293 ⣽⬊ࢆ⏝࠸࡚ࠊRNA-seq ゎᯒࢆ⾜ࡗ ࡓࠋ ەMed26 ࡢࣀࢵࢡ ࢘ࢺ࣐ ࢘ࢫࡼࡾ⥺ ⥔ⱆ⣽⬊ 㸦Med26-/-㸧ࢆᶞ❧ࡋࠊ㔝⏕ᆺࡢ࣐࢘ࢫ⥺⥔ⱆ⣽⬊ Med26-/-࣐࢘ࢫ⥺⥔ⱆ⣽⬊ࢆ⏝࠸࡚ RNA-seq ゎ ᯒࢆ⾜ࡗࡓࠋ ەMed26 ࡢ NTD㸦1-113㸧ࡢ 61 ␒┠ࡢ R㸦Arg㸧 62 ␒┠ࡢ K㸦Lys㸧ࢆ A㸦Ala㸧⨨ࡋࡓኚ␗ᆺ 㸦Med26-mut㸧ࡣࠊSEC ࡸ LEC ࡢ⤖ྜ⬟ࢆኻ࠺ࠋ ࡑࡇ࡛ࠊMed26-/-࣐࢘ࢫ⥺⥔ⱆ⣽⬊ࢆ⏝࠸࡚ࠊ㔝⏕ ᆺ Med26 ኚ␗ᆺ Med26 ࢆࡑࢀࡒࢀᏳᐃⓎ⌧ࡍ ࡿ ⣽ ⬊ ᰴ ࢆ ᶞ ❧ ࡋ ࡓ 㸦 Med26-/- + Med26-wt ࠊ Med26-/- + Med26-mut㸧ࠋࡇࢀࡽࡢ⣽⬊ࢆ⏝࠸࡚ࠊ RNA-seq ゎᯒࢆ⾜࠸ࠊMed26-/- + Med26-mut ⣽⬊ ࠾࠸࡚㌿ࡀᢚไࡉࢀࡿ㑇ఏᏊࡢ⥙⨶ⓗ࡞ゎᯒࢆ ⾜ࡗࡓࠋ RASGRP2 㸦 RAS guanyl-releasing protein 2 㸧ࡸ Wnt6 㸦 Wingless-type MMTV integration site family, member 6㸧࡞ࡢ㑇ఏᏊࡢⓎ⌧ࡀࠊኚ␗ᆺ Med26 ࢆⓎ⌧ࡍࡿ⣽⬊ẚࠊ㔝⏕ᆺࡢ Med26 ࢆ Ⓨ⌧ࡍࡿ⣽⬊࠾࠸࡚ࠊ≉␗ⓗୖ᪼ࡍࡿࡇࡀุ ᫂ࡋࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡣ⭘⒆ࡢቑṪࡶ㛵㐃ࡋ࡚ ࠸ࡿࡇࡽࠊMed26 ࡣ⭘⒆㛵㐃㑇ఏᏊ㡿ᇦ SEC ࡸ LEC ࢆࣜࢡ࣮ࣝࢺࡍࡿࡇࡼࡗ࡚ࠊࡑࢀࡽࡢ㑇 ఏᏊ㡿ᇦ࡛ Pol II ࡢ୍Ṇࢆゎ㝖ࡋࠊ⭘⒆ࡢቑṪ ᐤࡋ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 Histone 㑇ఏᏊࡢ mRNA ࡣࠊ㏻ᖖࡣ࣏ࣜࢹࢽࣝ 㸦PolyA ຍ㸧ࡉࢀ࡞࠸ࠋࡇࢁࡀࠊᮏゎᯒࡼࡗ ࡚ࠊ Med26 ࢆࣀࢵ ࢡࢲ࢘ࣥ ࡍࡿࡇ ࡼࡗ࡚ࠊ Histone 㑇ఏᏊࡢ mRNA ࡀ࣏ࣜࢹࢽࣝࡉࢀࡿࡇ ࡀࢃࡗࡓࠋࡇࡢࡇࡽࠊMed26 ࢆࣀࢵࢡࢲ࢘ ࣥࡍࡿࡇࡼࡾࠊRNA-seq ゎᯒ࠾ࡅࡿ Histone 㑇ఏᏊ⩌ࡢⓎ⌧ࡀࠊぢࡅୖࠊቑຍࡋ࡚࠸ࡓࡇࡀ ࢃࡗࡓࠋࡇࡢ⤖ᯝࡣࠊMed26 ࢆྵࡴ࣓ࢹ࢚࣮ࢱ ࣮」ྜయࡀ Histone 㑇ఏᏊ⩌ࡢ㌿⤊⤖ࡢไᚚᶵ ⬟ࡋ࡚࠸ࡿࡇࢆ♧၀ࡋ࡚࠾ࡾࠊᚋࡢゎᯒࡀ㔜せ ࡞ࡿࠋ 㸺ᨭᡂᯝබ⾲ㄽᩥࣜࢫࢺ㸼 Takahashi H., Takigawa I., Watanabe M., Anwar D., Shibata M., Tomomori-Sato C., Sato S., Ranjan A., Seidel CW., Tsukiyama T., Mizushima W., Hayashi M., Ohkawa Y., Conaway JW., Conaway RC., Hatakeyama S.: MED26 regulates the transcription of snRNA genes through the recruitment of little elongation complex. Nature Commun., 6, 5941, 2015. 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 Med26 ࢆ HEK293 ⣽⬊࡛ࣀࢵࢡࢲ࢘ࣥࡍࡿࡇ ࡼࡗ࡚ࠊc-Myc ࡸ Hsp70 ࡢࡳ࡞ࡽࡎࠊc-Jun ࡞ࢆ ྵࡴࡉࡲࡊࡲ࡞⭘⒆㛵㐃㑇ఏᏊࡢⓎ⌧ࡀపୗࡍࡿࡇ ࡀࢃࡗࡓࠋࡇࡢࡇࡽࠊMed26 ࡣ⭘⒆㛵㐃 ࡍࡿ㑇ఏᏊࡢⓎ⌧ࢆไᚚࡍࡿࡇࡀࢃࡗࡓࠋࡲࡓࠊ ⯆῝࠸ࡇࠊHistone 㑇ఏᏊ࡞ࡢ୍㒊ࡢ㑇ఏ Ꮚ⩌࠾࠸࡚ࠊMed26 ࡢࣀࢵࢡࢲ࢘ࣥࡼࡗ࡚ࠊࡑ ࡢⓎ⌧ࡀቑຍࡍࡿࡇࡶุ᫂ࡋࡓࠋ ࡲࡓࠊ࣐࢘ࢫ⥺⥔ⱆ⣽⬊ᰴ㸦Med26-/- + Med26-wtࠊ Med26-/- + Med26-mut㸧ࢆ⏝࠸ࡓゎᯒࡼࡗ࡚ࠊ - 405 - බເᨭㄢ㢟㸸2015 ᖺᗘ Pol2 ࡢ㌿ఙ㛗࣭⤊⤖࣭ࣜࢧࢡࣝ㐣⛬࠾ࡅࡿࢳ࢙ࢵࢡ࣏ࣥࢺไᚚᶵᵓࡢゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ 㧗ᶫ ⚽ᑦ࣭ᾏ㐨ᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ㏆ᖺࡢࢤࣀ࣒࣡ࢻ࡞ゎᯒࡼࡾࠊ㠀ᖖከࡃࡢࣄ ࢺࡢ㑇ఏᏊ㸦⣙ 30㸣♧၀ࡉࢀࡿ㸧࡛ࠊ㌿㛤ጞ┤ ᚋ RNA ࣏࣓࣮ࣜࣛࢮ II㸦Pol II㸧ࡀࣉ࣮ࣟࣔࢱ࣮ ㏆ഐ㸦㌿㛤ጞⅬࡢ 20㹼50 ሷᇶୗὶ㸧୍࡛Ṇࡋ ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡾࠊ㑇ఏᏊⓎ⌧ࡢไᚚ࡛㌿ ఙ㛗ࡢࣉࣟࢭࢫࡀ㔜せ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿࡇ ࡀࢃࡗ࡚ࡁࡓࠋPol II ࡢ୍Ṇࡣཎࡀࢇ㑇ఏᏊ c-Myc ࢆጞࡵࠊⓎ⌧ࡢ㎿㏿࠶ࡿ࠸ࡣྠㄪࡋࡓㄏᑟࡀ ᚲせ࡞⇕ࢩࣙࢵࢡ㑇ఏᏊ Hsp70ࠊ⾑Ύᛂ⟅ᛶ㑇ఏᏊ ࡸⓎ⏕ไᚚ㑇ఏᏊ࡞ࠊከࡃࡢࣄࢺ㑇ఏᏊ࠾࠸࡚ ࡳࡽࢀࡿࠋPol II ࡢ୍Ṇࡀゎ㝖ࡉࢀ Pol II ࡀ᪂ ⏕ RNA ࡢྜᡂࢆ㛤ࡍࡿࡓࡵࡣ㌿ఙ㛗ᅉᏊࡢ ാࡁࡀᚲせ࡛࠶ࡿࠋࡇࢁࡀࠊࡇࢀࡽࡢ㌿ఙ㛗ᅉ Ꮚࡀࠊࡢࡼ࠺ࡋ࡚≉ᐃࡢ㑇ఏᏊ㡿ᇦᮇ≉␗ ⓗࣜࢡ࣮ࣝࢺࡉࢀࡿࡢࡘ࠸࡚ᮍ▱࡛࠶ࡗࡓࠋ ⏦ㄳ⪅ࡣࡇࢀࡲ࡛ࠊ࣓ࢹ࢚࣮ࢱ࣮」ྜయࡢࢧࣈ ࣘࢽࢵࢺ Med26 ࡀࠊ2 ࡘࡢ␗࡞ࡿ㌿ఙ㛗ᅉᏊ」ྜ య͇ Super elongation complex͇ 㸦SEC㸧͇ Little elongation complex͇ 㸦LEC㸧ࢆࠊࡑࢀࡒࢀ␗࡞ࡿ✀ 㢮ࡢ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡋࠊࡑࢀࡽࡢ㑇ఏᏊࡢ ㌿ఙ㛗ࢆಁ㐍ࡍࡿࡇࢆ᫂ࡽࡋࡓࠋࡇࢀࡲ࡛ ࡢゎᯒࡽࠊMed26 ࡣ SEC ࢆ c-Myc ࡸ Hsp70 㑇ఏ Ꮚ࡞ࡢࢱࣥࣃࢡ㉁ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊ㡿ᇦࠊ LEC ࢆ small nuclear RNA 㑇 ఏ Ꮚ ࡞ ࡢ non-coding RNA 㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡋࠊࡑࢀࡽ ࡢ㑇ఏᏊࡢ㌿ఙ㛗ࢆಁ㐍ࡍࡿࡇࡀࢃࡗࡓ ࠙Takahashi H, et al. Cell 2011ࠚ࠙Takahashi H, et al. Nature Communications 2015ࠚࠋ ᮏ◊✲࡛ࡣࠊMed26 ࡀ SEC ࡸ LEC ࢆࡢࡼ࠺࡞ 㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡍࡿࡢࡘ࠸࡚⥙⨶ⓗ ᫂ࡽࡍࡿࠋࡉࡽࠊMed26 ࡀࡢࡼ࠺࡞࣓࢝ࢽ ࢬ࣒ࡼࡗ࡚ࠊSEC LEC ࢆࡑࢀࡒࢀ␗࡞ࡿ✀㢮 ࡢ㑇ఏᏊ㡿ᇦࣜࢡ࣮ࣝࢺࡍࡿࡢࡘ࠸࡚᫂ࡽ ࡍࡿࠋ RNAࠊ࠶ࡿ࠸ࡣᮍ▱ࡢ non-coding RNA ⤖ྜࡍࡿ ࡇࡼࡗ࡚ࠊᶆⓗ㑇ఏᏊ㡿ᇦ≉␗ⓗࣜࢡ࣮ࣝ ࢺࡉࢀࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࠋࡑࡇ࡛ࠊLEC ࡢࢧࣈ ࣘࢽࢵࢺࡢ KIAA0947 ࡸ NARG2 ࡢ RIP㸦RNA immunoprecipitation 㸧 ࡸ CLIP 㸦 cross-linking immunoprecipitation㸧ࢩ࣮ࢡ࢚ࣥࢫゎᯒࢆ⾜࠸ࠊ LEC ⤖ྜࡍࡿ RNA ࡢ⥙⨶ⓗྠᐃࢆヨࡳࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏ◊✲ࡢ㐙⾜ࡼࡾࠊMed26 SECࠊ࠶ࡿ࠸ࡣ Med26 LEC ࡼࡗ࡚ࡑࢀࡒࢀไᚚࡉࢀࡿᶆⓗ㑇 ఏᏊࡀࠊࡉࡽ⥙⨶ⓗゎ᫂ࡉࢀࡿࡇࡀᮇᚅ࡛ࡁ ࡿࠋࡲࡓࠊLEC RNA ࡢ⤖ྜࡀࠊMed26 ࡼࡿ LEC ࡢࣜࢡ࣮ࣝࢺᶵᵓ㛵ࡋ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࢀࡤࠊMed26 ࡼࡿ SEC ࡸ LEC ࡢᶆⓗ㑇ఏᏊ Ỵᐃᶵᵓࡢ୍➃ࡀ᫂ࡽ࡞ࡿࡇࡶᮇᚅ࡛ࡁࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ๓ᖺᗘࡽࡢᨭࡼࡗ࡚ࠊMed26 ࡣ୍㒊ࡢ㑇ఏᏊ 㸦mRNA ࡀ࣏ࣜࢹࢽࣝࡉࢀ࡞࠸㑇ఏᏊ㸧ࡢ㌿ ⤊⤖ࡢไᚚᶵ⬟ࡋ࡚࠸ࡿࡇࡀ♧၀ࡉࢀࡓࠋᮏ◊ ✲ࡢ᥎㐍ࡼࡾࠊMed26 ࡀ SEC ࡸ LEC ࢆ㑇ఏᏊ㡿 ᇦࣜࢡ࣮ࣝࢺࡍࡿࡇࡼࡗ࡚ࠊ㌿ఙ㛗ࡢࡳ࡞ ࡽࡎ㌿⤊⤖ࡢไᚚࡶᶵ⬟ࡍࡿࡇࡀ᫂ࡽ࡞ ࡿྍ⬟ᛶࡀ࠶ࡿࠋ࣓ࢹ࢚࣮ࢱ࣮」ྜయࡢ㌿⤊⤖ ࠾ࡅࡿᶵ⬟ࡣࢇᮍ▱࡛࠶ࡿࡇࡽࠊᮏ◊ ✲ࡢ㐙⾜ࡣࠊ㑇ఏᏊⓎ⌧ไᚚࡢ◊✲࠾࠸࡚ࡶࠊ1 ࡘࡢࣈ࣮ࣞࢡࢫ࣮ࣝ࡞ࡿࡇࡀᮇᚅ࡛ࡁࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾࠊㄽᩥࡢᢞ✏‽ഛࢆ⾜ࡗ࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ەMed26ࠊSECࠊLEC ࡼࡗ࡚Ⓨ⌧ࡀไᚚࡉࢀࡿ㑇 ఏᏊࢆ⥙⨶ⓗ᫂ࡽࡍࡿࡓࡵࠊMed26ࠊAFF4 㸦SEC ࡢࢧࣈࣘࢽࢵࢺ㸧ࠊKIAA0947㸦LEC ࡢࢧࣈ ࣘࢽࢵࢺ㸧ࢆࡑࢀࡒࢀࣀࢵࢡࢲ࢘ࣥࡋࡓ HEK293 ⣽ ⬊ࢆ⏝࠸࡚ RNA-seq ゎᯒࢆ⾜࠺ࠋ ەLEC ࡼࡗ࡚Ⓨ⌧ࡀไᚚࡉࢀࡿ㑇ఏᏊࢆࡉࡽ⥙ ⨶ⓗ᫂ࡽࡍࡿࡓࡵࠊMicrococcal nuclease ࢆ⏝࠸ࡓ ChIP ἲࡼࡾࠊLEC ࢧࣈࣘࢽࢵࢺࡢ KIAA0947 ࡸ NARG2 ࡢ ChIP-seq ゎᯒࢆ⾜࠺ࠋ ەLEC ࡢࢧࣈࣘࢽࢵࢺࡢ NARG2 ࡣ RNA ࡢ⤖ ྜᶵ⬟ࡍࡿணࡉࢀࡿࢻ࣓ࣥࡀᏑᅾࡍࡿࠋࡇ ࡢࡇࡽࠊLEC ࡣᶆⓗ㑇ఏᏊࡼࡾ㌿ࡉࢀࡿ᪂⏕ - 406 - බເᨭㄢ㢟㸸2014 ᖺᗘ 㝣ୖ᳜≀ࡢỈ⋓ᚓᶵ⬟ࡍࡿ᰿ࡢỈᛂ⟅ᶵᵓࡢゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ 㧗ᶫ ⚽ᖾ࣭ᮾᏛᏛ㝔⏕⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 1930 ᖺ௦ᥦၐࡉࢀࡓࢥࣟࢻࢽ࣮࣭࢙࢘ࣥࢺㄝࡣ᳜ ≀⛉Ꮫࡢせ࡞⌮ㄽࡢ୍ࡘ࡛࠶ࡿࠋᮏ௬ㄝ࡛ࡣࠊᒅ ᛶࢆⓎ⌧ࡍࡿჾᐁࡣࠊ⎔ቃ่⃭ᛂ⟅ࡋࡓ࣮࢜࢟ࢩ ࣥ㍺㏦ࡢኚࡼࡾࠊ࣮࢜࢟ࢩࣥࡢ೫ᕪศᕸࡀㄏᑟ ࡉࢀࠊ೫ᕪᡂ㛗ࡀㄏᑟࡉࢀࠊᒅ᭤ࡀ⏕ࡌࡿࠊᒅᛶ ࢆㄝ᫂ࡋ࡚࠸ࡿࠋ⏦ㄳ⪅ࡣ⌧ᅾࡲ࡛ࠊỈศᒅᛶࡢ Ⓨ⌧࣮࢜࢟ࢩࣥ㍺㏦ࢆᚲせࡍࡿ᳜≀✀ (ࢿࠊ ࢟ࣗ࢘ࣜࠊ࢚ࣥࢻ࢘) ࠊ࣮࢜࢟ࢩࣥ㍺㏦ࢆᚲせ ࡋ࡞࠸᳜≀✀ (ࢩࣟࢾࢼࢬࢼࠊ࣑ࣖࢥࢢࢧ) ࡀ࠶ࡿ ࡇࢆぢฟࡋࡓࠋࡑࡋ࡚ࠊࡇࢀࡽ࡚ࡢ᳜≀✀ࡢ㔜 ຊᒅᛶࡣ࣮࢜࢟ࢩࣥ㍺㏦ࢆᚲせࡍࡿࡇࢆ☜ㄆࡋ ࡓࠋᮏ◊✲࡛ࡣỈศᒅᛶࡢ≉ᛶࡀ␗࡞ࡿ᳜≀✀ࡢỈ ศᒅᛶ㔜ຊᒅᛶࡢ㑇ఏᏊⓎ⌧ࢆ⥙⨶ⓗゎᯒ ͌ẚ㍑ࡋࠊ೫ᕪⓗⓎ⌧ࡍࡿ㑇ఏᏊ⩌ࡢရ✀㛫࡛ࡢᕪ ␗ࠊ࠾ࡼࡧࠊྠ୍᳜≀✀㛫ᒅᛶ㛫࡛ࡢᕪ␗ࢆぢฟࡍ ࡇࢆ┠ⓗࡋ࡚ࠊゎᯒࢆ⾜ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 㟼⨨᮲௳ୗ࡛ࡶ᰿ࡢỈศᒅᛶࢆⓎ⌧ࡋ (Ỉศᒅᛶ> 㔜ຊᒅᛶ)ࠊ࣮࢜࢟ࢩࣥ㍺㏦㜼ᐖឤཷᛶỈศᒅᛶࢆ Ⓨ⌧ࡍࡿ᳜≀✀ࡋ࡚ࠊࢿ (Oryza sativa) ࢆࠊ㟼 ⨨᮲௳ୗ࡛ࡶ᰿ࡢỈศᒅᛶࢆⓎ⌧ࡋ (Ỉศᒅᛶ>㔜 ຊᒅᛶ)ࠊ࣮࢜࢟ࢩࣥ㍺㏦㜼ᐖ⪏ᛶỈศᒅᛶࢆⓎ⌧ ࡍࡿ᳜≀✀ࡋ࡚ࠊ࣑ࣖࢥࢢࢧ (Lotus japonicas) ࢆࠊࡑࡋ࡚ࠊ㟼⨨᮲௳ୗ࡛ࡣỈศᒅᛶࢆⓎ⌧࡛ࡁࡎࠊ ࢡࣜࣀࢫࢱࢵࢺ᮲௳ୗ࡛ỈศᒅᛶࢆⓎ⌧ࡋ (Ỉศᒅ ᛶ<㔜ຊᒅᛶ)ࠊ࣮࢜࢟ࢩࣥ㍺㏦㜼ᐖឤཷᛶỈศᒅ ᛶࢆⓎ⌧ࡍࡿ᳜≀✀ࡋ࡚ࠊ࢟ࣗ࢘ࣜ (Cucumis sativus) ࢆ⏝࠸ࡓࠋࡑࡋ࡚ࠊỈศᒅᛶࢆⓎ⌧ࡉࡏ࡚ ࠸ࡿࢿ࣑ࣖࢥࢢࢧࡢ᰿ࡢ㑇ఏᏊⓎ⌧ࢆゎᯒࡍࡿ ࡓࡵࠊᐮኳᇵᆅୖ࡛⏕⫱ࡉࡏࡓⱆ⏕࠼ࢆ⏝࠸ࠊỈศ ໙㓄่⃭ࢆ࠼ࠊ⏕⫱ࡉࡏࡓⱆ⏕࠼ࡢࠊ㧗Ỉศഃ (ᐮ ኳഃ) పỈศഃ (ᐮኳഃ) ษࡾศࡅࡓ᰿ࠊࢆࡑ ࢀࡒࢀࢧࣥࣉࣜࣥࢢࡋࠊRNA ࢆᢳฟࡋࡓࠋ ࡲࡓࠊ㔜ຊᒅᛶࢆⓎ⌧ࡋ࡚࠸ࡿࢿࠊ࣑ࣖࢥࢢࢧࠊ ࢟ࣗ࢘ࣜࡢ᰿ࡢ㑇ఏᏊⓎ⌧ࢆゎᯒࡍࡿࡓࡵࠊࢿ ࣑ࣖࢥࢢࢧࡣᆶ┤⨨࠸ࡓᐮኳᇵᆅୖ࡛⏕⫱ࡉࡏࡓ ⱆ⏕࠼ࢆ⏝࠸ࠊ࢟ࣗ࢘ࣜࡣỈศ྾ᛶࡢࣉࣛࢫࢳࢵ ࢡ✀Ꮚࢆᕪࡋ㎸ࢇ࡛ᆶ┤⏕⫱ࡉࡏࡓⱆ⏕࠼ࢆ⏝ ࠸ࠊⱆ⏕࠼ࢆᶓಽࡋ࡚ࠊ㔜ຊ่⃭ࢆ࠼ࠊ⏕⫱ࡉࡏ ࡓⱆ⏕࠼ࡢୖୗษࡾศࡅࡓ᰿ࠊࢆࡑࢀࡒࢀࢧࣥࣉ ࣜࣥࢢࡋࠊRNA ࢆᢳฟࡋࡓࠋ ࡇࢀࡽࡢ RNA ࡽࡢ Oligo-dT ࣅ࣮ࢬࢆ⏝࠸ࡓ poly(A) RNA ࡢ⢭〇ࠊpoly(A)-RNA ࡢ᩿∦ࠊcDNA ࡢྜᡂࠊྛ᳨య㆑ู⏝ࡢࣥࢹࢵࢡࢫࢱࢢࢆྵࡴ ࢲࣉࢱ࣮ࡢࣛࢤ࣮ࢩࣙࣥࠊIllumina HiseqTM 2500 ࡼࡿࢩ࣮ࢡ࢚ࣥࢫࠊᚓࡽࢀࡓሷᇶ㓄ิ 㸦FASTAQ ࣇࣝ㸧 ࡢ TophHat㸦2.1.10㸧ࡼࡿࢤࣀ࣒ሷᇶ 㓄ิࡢ࣐ࢵࣆࣥࢢࠊcuffdiff ࡼࡿⓎ⌧ゎᯒࢆᨭ ࡋ࡚㡬࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࢟ࣗ࢘ࣜࡢ㔜ຊᒅᛶࢆⓎ⌧ࡉࡏࡓ᰿ࡢୖഃୗഃ࡛ ࡢⓎ⌧ࡢᕪ␗ࡀࠊࡢ㑇ఏᏊẚ㍑ࡋ࡚᭷ពㄆࡵ ࡽࢀࡿ 38 㑇ఏᏊࢆぢฟࡋࡓࠋࡇࢀࡽࡢ࠺ࡕࠊ19 㑇 ఏᏊࡀୗഃ࡛㧗Ⓨ⌧ࡋࠊ࢜ࣥࢺࣟࢪ࣮ゎᯒࡢ⤖ᯝࠊ ࣮࢜࢟ࢩࣥㄏᑟᛶ㑇ఏᏊ⩌ࡀࠊ㔜ຊᒅᛶ᰿ࡢୗ ഃ࡛Ⓨ⌧ࡋ࡚࠸ࡿࡇࡀ♧ࡉࢀࡓࠋୗഃ࡛㧗Ⓨ⌧ࡍ ࡿ 19 㑇ఏᏊࡣࠊ࢜ࣥࢺࣟࢪ࣮ゎᯒࡋࡓ⤖ᯝࠊ≉ᐃࡢ 㑇ఏᏊ⩌ᒓࡋ࡚࠸࡞ࡗࡓࠋࡋࡓࡀࡗ࡚ࠊ࢟ࣗ࢘ ࣜࡢ᰿ࡢ㔜ຊᒅᛶࢆᘬࡁ㉳ࡇࡍせᅉࡣࠊ᰿ࡢୗഃ ✚ࡍࡿ࣮࢜࢟ࢩ࡛ࣥ࠶ࡿࡇࡀࠊᮏゎᯒࡽࡶ ᨭᣢࡉࢀࡓࠋ ୍᪉ࠊࢿࡢ㔜ຊᒅᛶࢆⓎ⌧ࡉࡏࡓ᰿ࡢୖഃୗ ഃࠊ࠾ࡼࡧỈศᒅᛶࢆⓎ⌧ࡉࡏࡓ᰿ࡢ㧗Ỉศഃప Ỉศഃ࡛ࡢⓎ⌧ࡢᕪ␗ࡀࠊࡢ㑇ఏᏊẚ㍑ࡋ࡚᭷ ពㄆࡵࡽࢀࡿ㑇ఏᏊ (q < 0.05) ࡣぢฟࡏ࡞ࡗࡓࠋ ࡲࡓࠊ࣑ࣖࢥࢢࢧࡢ㔜ຊᒅᛶࢆⓎ⌧ࡉࡏࡓ᰿ࡢୖഃ ୗഃ࡛ࡢⓎ⌧ࡀࠊࡢ㑇ఏᏊẚ㍑ࡋ࡚᭷ពᕪ ␗ࡀ࠶ࡿ♧ࡉࢀࡓ 42 㑇ఏᏊ (q < 0.05) ୰ 3 㑇ఏ Ꮚࡀࣜ࣎ࢯ࣮࣒ RNA ࡛࠶ࡾࠊࡲࡓࠊỈศᒅᛶࢆⓎ⌧ ࡉࡏࡓ᰿ࡢ㧗ỈศഃపỈศഃ࡛ࡢⓎ⌧ࡢᕪ␗ࡀࠊ ࡢ㑇ఏᏊẚ㍑ࡋ࡚᭷ពᕪ␗ࡀ࠶ࡿ♧ࡉࢀࡓ 29 㑇ఏᏊ (q < 0.05) ୰ 15 㑇ఏᏊࡀࣜ࣎ࢯ࣮࣒ RNAࠊ ࠶ࡿ࠸ࡣࣜ࣎ࢯ࣮࣒ࢱࣥࣃࢡ㉁ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊ ࡔࡗࡓࠋࡋࡓࡀࡗ࡚ࠊᅇࡢࢿ࣑ࣖࢥࢢࢧࡢ RNA-Seq ࡣࠊmRNA ࡢ✚ࢆṇ☜᳨ฟ࡛ࡁࡓ ࡣゝ࠸ࡀࡓ࠸⤖ᯝ࡛࠶ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ⌧ᅾࠊ࢟ࣗ࢘ࣜࡢ᰿ࡢỈศᒅᛶ೫ᕪⓗⓎ⌧ࢆࡍ ࡿ㑇ఏᏊࢆ RNA-Seq ࡼࡾゎᯒࡋࡓ⤖ᯝࠊᨭ ࡼࡗ࡚ᚓࡽࢀࡓゎᯒ⤖ᯝࢆẚ㍑ࡋ࡚࠸ࡿࠋᮏẚ㍑ ゎᯒࡼࡾࠊ࢟ࣗ࢘ࣜࡢ᰿ࡢ㔜ຊᒅᛶỈศᒅᛶࡢ Ⓨ⌧ᶵᵓࡢ㢮ఝᛶ┦㐪ᛶࡀ᫂ࡽ࡞ࡿᮇᚅࡉ ࢀࡿࠋࡲࡓࠊᅇࠊRNA-Seq ࡢゎᯒ⤖ᯝࡀᛮࢃࡋࡃ ࡞ࡗࡓࢿ࣑ࣖࢥࢢࢧࡘ࠸࡚ࡣࠊᒅᛶࢆⓎ⌧ ࡉࡏࡿᐇ㦂᪉ἲࠊ࠾ࡼࡧゎᯒ⏝࠸ࡿ᳜≀✀ࡢ㑅ᐃ ࢆྵࡵࠊ᳨ウࡋࠊᮏ◊✲ㄢ㢟ࡢỈศᒅᛶࡢⓎ⌧ᶵ ᵓࡢ᳜≀✀㛫ᕪࡀ⏕ࡌࡿᶵᵓࡢゎ᫂ࢆ┠ᣦࡋࠊ◊✲ ࢆᒎ㛤ࡋ࡚ࡁࡓ࠸⪃࠼࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ㄽᩥࣜࢫࢺ㸼 ࡞ࡋ - 407 - බເᨭㄢ㢟㸸2014 ᖺᗘ 〓⸴ࡢ㟷Ⰽගᛂ⟅ᛂࡢゎᯒ ᇶ┙◊✲㸦㹁㸧 㧗ᶫ ᩥ㞝࣭❧㤋Ꮫ⏕⛉Ꮫ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 2007 ᖺࠊ᪂ወࡢ㌿ᅉᏊᆺ㟷Ⰽගཷᐜయࠊ࣮࢜ ࣞ࢜ࢡ࣒ࣟ㸦AUREOCHROME㸧ࢆⓎぢࡋࡓࠋ࣮࢜ ࣞ࢜ࢡ࣒ࣟࡣࠊ〓⸴ࡸࢣ⸴ࢆྵࡴ㯤Ⰽ᳜≀㸦⊂❧ ᰤ㣴ࢫࢺ࣓ࣛࣀࣃࣝ㸧ࡢࡳࡼࡃಖᏑࡉࢀ࡚࠾ࡾࠊ 㯤Ⰽ᳜≀ඹ㏻ࡢ㔜せ࡞ᶵ⬟ࢆᢸࡗ࡚࠸ࡿ᥎ᐃ࡛ࡁ ࡿࠋ࣮࢜ࣞ࢜ࢡ࣒ࣟࡣ㯤Ⰽ᳜≀ᒓࡍࡿ㯤⥳⸴ࣇࢩ ࢼࢩ࣑ࢻ࡛ࣟࡣ㟷Ⰽගㄏᑟศᯞᙧᡂ㛵ࡋ࡚࠸ࡿ ࡀࠊ〓⸴ࡸࢣ⸴ࡣࡌࡵࡢ㯤Ⰽ᳜≀࡛ࡢᶵ⬟ࡣ ࡃᮍゎ࡛᫂࠶ࡿࠋᮏ◊✲ࡢ┠ⓗࡣࠊᐊෆᐇ㦂࡛ᇵ㣴 ⣔ࢆ☜❧ࡋࡓ〓⸴ࣖࣁࢬࢢࢧ㸦Dictyopteris㸧࢝ ࣔࢡ㸦Sargassum㸧ࢆ⏝࠸࡚ࠊࡢࡼ࠺࡞ගᛂࡀ ࣮࢜ࣞ࢜ࢡ࣒ࣟࡼࡗ࡚ไᚚࡉࢀ࡚࠸ࡿࢆ᫂ࡽ ࡋࠊ㯤Ⰽ᳜≀ඹ㏻ࡢᶆⓗᛂࢆ᥎ᐃࡍࡿࡇ࡛࠶ ࡿࠋ 㸺ᨭࡢෆᐜ㸼 RNAseq㸦ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒㸧ゎᯒ ගࡣࠊ᳜≀㸦┿᰾ගྜᡂ⏕≀㸧ࡗ࡚ࠊගྜᡂࡔ ࡅ࡛࡞ࡃගཷᐜయࢆࡋࡓಙྕࡋ࡚⏝࠸ࡽࢀࠊᙧ ែᙧᡂᛂࡸ㐠ືᛂࢆᘬࡁ㉳ࡇࡍࠋ㝣ୖ᳜≀ࢆྵ ࡴ⥳Ⰽ᳜≀࡛ࡣࠊ✺↛ኚ␗యࡸᙧ㉁㌿య᳜≀ࢆ ⏝ࡋࠊつᶍ࡞ࢤࣀ࣒ゎᯒࡸࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ ゎᯒࡀ⾜ࢃࢀࠊࡑࡢගཷᐜయࡢㇺࡀ᫂ࡽ࡞ࡗ ࡚ࡁࡓࠋࡋࡋࠊỈᇦ⦾ⱱࡋࠊ㐍࣭⣔⤫ⓗ ࡁࡃ␗࡞ࡿ㯤Ⰽ᳜≀ࡣࠊ⥳Ⰽ᳜≀ࡢࡼ࠺࡞ࢤࣀ࣒ࢹ ࣮ࢱ࣮࣋ࢫࡣᑡ࡞ࡃࠊගཷᐜయࡢᏑᅾᑐࡍࡿࢹ࣮ ࢱࡣஈࡋ࠸ࠋᮏ⏦ㄳ࡛ࡣࠊྂࡃࡽගᙧែᙧᡂᛂ ࡀほᐹࡉࢀࠊᐇ㦂ᐊෆ࡛ᇵ㣴ࡀྍ⬟࡞ࡗࡓ㯤Ⰽ᳜ ≀ᒓࡍࡿ〓⸴㢮㸦᪻ᕸࡸ࣓࣡࢝ࡢ௰㛫㸧ࢆ⏝࠸ࠊ mRNA ࡢつᶍゎᯒࢆ⾜࠸ࠊගཷᐜయࡢ᥈⣴ࢆ࡞ ┠ⓗࡍࡿࠋලయⓗ࡞ᮦᩱࡣࢤࣀ࣒ሗࡀஈࡋ࠸〓 ⸴ࣄࣂ࣐ࢱ⛉㸦ࢤࣀ࣒ࢧࢬ⣙ 1000 Mb㸧ᒓࡍࡿ ࢝ࣔࢡࡢᰤ㣴⸴య࡛Ⓨ⌧ࡋ࡚࠸ࡿ mRNA ࢆࠊḟୡ ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮ࢆ⏝࠸࡚ゎㄞࡍࡿࡇࡼࡾࠊ ࢝ࣔࢡࢤࣀ࣒ྵࡲࢀࡿගཷᐜయ㑇ఏᏊ⩌せ㑇 ఏᏊࡢ࢝ࢱࣟࢢࢆ┠ⓗࡍࡿࠋࡇࡢࢹ࣮ࢱࢆࡶ ࠊᵝࠎ࡞ගᇵ㣴᮲௳ୗࡢ㑇ఏᏊࡢⓎ⌧ᶵᵓࢆㄪ ࡿࠋ ࣒ࡢศᏊࡀྛ 5 㑇ఏᏊࠊ2 㑇ఏᏊ࠶ࡿࡇࡀࢃࡗ ࡓࠋ୍᪉࡛ 4 ࢧࣥࣉࣝ㛫ࡢ㑇ఏᏊࡢቑῶࢆㄪࡓ ࡇࢁࠊ⯆῝࠸ࡇ myb-like ࡢ㌿ᅉᏊࡢⓎ⌧ࡢ ᕪࡀ㢧ⴭ⾲ࢀࡿࡇࡀศࡗࡓࠋࡲࡓࡢ㌿ᅉ Ꮚ⩌㸦c6-type㸧ࡢ㑇ఏᏊ⩌ࡶⓎ⌧ᕪࡀ࠶ࡿࡇࡶศ ࡗࡓࠋ⌧ᅾ real time PCR ࢆ⾜࠸ࠊ㏆᪥බ㛤࣭ㄽ ᩥసᡂࢆணᐃࡋ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᚓࡽࢀࡓࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢹ࣮ࢱ RT-PCR ࡢ」ྜἲ࡛㔜せ࡞㑇ఏᏊࡢ㛗༢㞳ࡶ⾜ࡗ࡚࠸ࡿࠋ ࡑࢀࡽࡀ᪩ᮇᐇ⌧ࡍࢀࡤࠊ⛉◊㈝⏦ㄳࡢෆᐜ࡛࠶ ࡿࠊගཷᐜయࢱࣥࣃࢡ➼ࡢ⢭〇࣭ศගゎᯒ➼ࡢ◊✲ ࢆ㐍ࡵࡓ࠸⪃࠼࡚࠸ࡿࠋ ᨭࢆཷࡅࡓࡇࡼࡗ࡚ࠊᶵ⬟ⓗ࡞ࢱࣥࣃࢡ㉁ ࡢⓎ⌧ࡼࡾࠊ㌿ᅉᏊࢱࣥࣃࢡ㉁ࡢⓎ⌧ᕪࡀࡳࡽ ࢀࡓࡇࡽࠊ㑇ఏᏊୖὶࡢࣉ࣮ࣟࣔࢱ࣮㓄ิࡢྲྀ ᚓࢆヨࡳࡿࡇࡀᚲ㡲࡞ࡗࡓࠋ⌧ᅾࠊPacBio ࢆ⏝ ࠸ࡓࢤࣀ࣒ゎㄞࢆ⾜ࡗ࡚ࠊ㌿ᅉᏊ⩌ࡀ⤖ྜࡍࡿ㓄 ิྲྀᚓࢆヨࡳࡓࠋ⌧ᅾࠊྲྀᚓࡋࡓࢤࣀ࣒㓄ิࢆ⏝࠸ ࡚ࣉ࣮ࣟࣔࢱ࣮ゎᯒࢆ⾜ࡗ࡚࠸ࡿࠋ ᮏࢤࣀ࣒ᨭࡼࡗ࡚ࠊග᮲௳ୗ࡛Ⓨ⌧ࡢቑῶࡍ ࡿ㑇ఏᏊ⩌ࡀ㌿ᅉᏊࡀከᩘ࡛࠶ࡗࡓࡇࡽࠊࢤ ࣀ࣒ࢆゎㄞࢆヨࡳࡿ⮳ࡗࡓࠋୗὶࡢ㌿ᅉᏊ࢝ࢫࢣ ࣮ࢻࡀࠊ㯤Ⰽ᳜≀࡛ࡣࡌࡵ࡚ゎᯒࡉࢀࡓ࡞ࡿ ᛮࢃࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 㸺Ꮫ㸼 ྜྷᕝఙဢ㸦⚟┴࣭ᾏὒ㸧 ⣽ᕝ┿⨾㸦⚟┴࣭ ᾏὒ㸧 ୖ㐍ஓ㸦᪂₲࣭⌮㸧 ዟ⏣ಟ㑻㸦᪂ ₲࣭ṑ㸧 ➟ཎ㈼ὒ㸦❧㤋࣭⏕㸧 㧗ᶫᩥ㞝 㸦❧㤋࣭⏕㸧 RNA-seq ࡼࡿ〓⸴࢝ࣔࢡࡢ㟷Ⰽගཷᐜయࡢゎᯒ ᪥ᮏ⸴㢮Ꮫ 2016 ᖺ 3 ᭶ 19㸫20 ᪥ணᐃ ᪥ᮏṑ⛉Ꮫ㸦ᮾி㸧 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࢝ࣔࢡ㞝ᰴ 4 ࢧࣥࣉࣝ㸦ග᮲௳ࢆኚࡋࡓ㸧ࡢ ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ⾜࠸ࠊࢹ࣮ࢱ࣮࣋ࢫᵓ ⠏ࢆᖺᗘ୰⤊ࡉࡏࡓࠋࢭࣥࣈࣝࠊࣀࢸ࣮ ࢩࣙࣥ➼ࡣࠊ↛ࡿࡁ◊✲⪅ඹྠࡋ࡚⾜࠸ࠊ4 ࢧ ࣥ ࣉ ࣝ 㛫 ࡢ Ⓨ ⌧ 㔞 ࡢ ᕪ ࢆ ㄪ ᰝ ࡋ ࡓ 㸦 http://bioinfo.med.niigata-u.ac.jp/takahashi2/ ࠋ ࡲࡎ㟷Ⰽගཷᐜయ 8 㑇ఏᏊ᳨ฟࡋࠊᙧែᙧᡂ㛵 ࡍࡿ⪃࠼ࡽࢀࡿ࣮࢜ࣞ࢜ࢡ࣒ࣟࢡࣜࣉࢺࢡ࣮ࣟ - 408 - බເᨭㄢ㢟㸸2015 ᖺᗘ HP1 FACT ࡢඹᙺࡼࡿࢢ࣮ࣟࣂࣝ࡞㌿ไᚚᶵᵓ ᪂Ꮫ⾡㡿ᇦ◊✲ 㧗⏿ ಙஓ࣭ᾏ㐨ᏛᏛ㝔⌮Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 ࣊ࢸࣟࢡ࣐ࣟࢳࣥ㧗ḟᵓ㐀ࡣ୍㐣ⓗ㌿ࡉ ࢀࡿ㛗㙐㠀ࢥ࣮ࢻ RNA㸦lncRNA㸧ࢆᛂᇶ㉁ ࡍࡿ RNAi ౫Ꮡࡋ࡚☜❧ࡉࢀࡿ≉ᚩࢆᣢࡕࠊ ࠸ࡗࡓࢇ☜❧ࡉࢀࡓ࣊ࢸࣟࢡ࣐ࣟࢳࣥࡣ FACT ࢆྵࡴ」ᩘࡢ࢚ࣇ࢙ࢡࢱ࣮ࢱࣥࣃࢡ㉁ࡼࡗ࡚ Ᏻᐃ⥔ᣢࡉࢀࡿ࣮ࣔࢻ⛣⾜ࡍࡿࠋFACT ࡀ ࣊ࢸࣟࢡ࣐ࣟࢳࣥ㡿ᇦෆ࡛ࢧࣞࣥࢩࣥࢢ ᐤࡍࡿศᏊ࣓࢝ࢽࢬ࣒ࢆ㑇ఏᏛⓗ࣭⏕Ꮫⓗ ゎᯒࡋࡓ⤖ᯝࠊ࣊ࢸࣟࢡ࣐ࣟࢳࣥࢱࣥࣃࢡ㉁ HP1 FACT ࡀ≀⌮ⓗᙉࡃ┦స⏝ࡋ࡚ lncRNA ࡢ㌿ࢆไᚚࡋ࡚࣊ࢸࣟࢡ࣐ࣟࢳࣥࢆ Ᏻᐃ⥔ᣢࡍࡿࢆぢ࠸ࡔࡋࡓࠋ᭦⯆῝࠸ ⏦ㄳ⪅ࡽࡣ HP1 FACT ࡼࡿࢡ࣐ࣟࢳࣥไ ᚚ⣔ࡣ࣮ࣘࢡ࣐ࣟࢳࣥୖ࡛ࣄࢫࢺࣥ H3K9 ࡢ࣓ ࢳࣝ㸦H3K9me㸧㠀౫Ꮡⓗࡶ㉳ࡁࡿ⌧㇟࡛࠶ ࡿࢆⓎぢࡋ࡚࠾ࡾࠊᮏ◊✲ㄢ㢟࡛ࡣࡑࡢศᏊ ࣓࢝ࢽࢬ࣒⏕≀Ꮫⓗព⩏ࡢゎ᫂ࢆ┠ᣦࡍࠋ 㸺ᨭࡢෆᐜ㸼 ᮏ◊✲࡛ࡣࠊศ㓝ẕ࣊ࢸࣟࢡ࣐ࣟࢳࣥࢱࣥ ࣃࢡ㉁ HP1 ࡑࢀ≀⌮ⓗ⤖ྜࡋ࡚㑇ఏᏊⓎ ⌧ㄪ⠇㛵ࢃࡿ FACT 」ྜయࡢࢢ࣮ࣟࣂࣝ࡞ࢡ ࣐ࣟࢳࣥไᚚᶵᵓࢆ᥈ࡿࢆ┠ⓗࡋ࡚࠸ࡿࠋ ࡇࡢࢩࢫࢸ࣒ࡣ RNAi ᅉᏊ࣭ࢡ࣐ࣟࢳࣥࢱࣥࣃ ࢡ㉁࣭࣊ࢸࣟࢡ࣐ࣟࢳࣥࢱࣥࣃࢡ㉁ࡀ┦స⏝ ࡋࡘࡘࠊ࣊ࢸࣟࢡ࣐ࣟࢳࣥࡢࡳ࡞ࡽࡎࠊࢤࣀ࣒ ୖࡢከࡃࡢ㑇ఏᏊࡢ㌿ࢆไᚚࡋ࡚࠸ࡿࢆ ᐃࡋ࡚࠸ࡿⅭࠊ௨ୗࡢ㏻ࡾ &K,3íVHT RNA-seq ࢆィ⏬ࡋࡓࠋ ࣭࣊ࢸࣟࢡ࣐ࣟࢳࣥ㛵㐃ᅉᏊኚ␗ᰴࢆ⏝࠸ࡓ HP1 ศ 㓝ẕ࣍ࣔࣟࢢ Swi6 ࡢ ChIPѸseq ィ㸱ࢧࣥࣉࣝ ࣭࣊ࢸࣟࢡ࣐ࣟࢳࣥ㛵㐃ᅉᏊኚ␗ᰴࢆ⏝࠸ࡓ HP1 ศ 㓝ẕ࣍ࣔࣟࢢ Chp2 ࡢ ChIPѸseq ィ㸱ࢧࣥࣉࣝ ࣭࣊ࢸࣟࢡ࣐ࣟࢳࣥ㛵㐃ᅉᏊኚ␗ᰴࢆ⏝࠸ࡓ FACT ᵓᡂᅉᏊ Spt16 ࡢ ChIPѸseq ィ㸲ࢧࣥࣉࣝ ࣭࣊ࢸࣟࢡ࣐ࣟࢳࣥ㛵㐃ᅉᏊኚ␗ᰴࢆ⏝࠸ࡓ RNA-seq ィ㸰㸮ࢧࣥࣉࣝ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏ◊✲ࡣ࣊ࢸࣟࢡ࣐ࣟࢳࣥᙧᡂᅉᏊࡢ୰࡛ࡶ ≉୰ᚰⓗ࡞ᙺࢆᯝࡓࡍ࣊ࢸࣟࢡ࣐ࣟࢳࣥࢱ ࣥࣃࢡ㉁ࡑࡢ┦స⏝ᅉᏊ FACT 」ྜయ╔ ┠ࡋ࡚ゎᯒࢆࡍࡍࡵࡓࠋHP1/Swi6 ࡢ ChIPѸseq ゎᯒ⤖ᯝࡽࠊ࣊ࢸࣟࢡ࣐ࣟࢳࣥෆ࡛ࡣ HP1/Swi6 ࡀ࣊ࢸࣟࢡ࣐ࣟࢳࣥࡢ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࣐࣮ࢡ ࡋ࡚▱ࡽࢀࡿ H3K9me 㠀ᖖ㧗ࡃඹᒁᅾࡋ࡚࠸ ࡿࡀ᫂ࡽ࡞ࡗࡓࠋࡲࡓ H3K9me ࡢᾘኻక࠸ࠊ ࡇࡢ HP1/Swi6 ࡢ࣊ࢸࣟࢡ࣐ࣟࢳࣥࡢ⤖ྜࡶࡲࡓ ᾘኻࡍࡿࡀ᫂ࡽ࡞ࡗࡓࠋྠ࣮ࣘࢡ࣐ࣟࢳ ࣥୖ࡛ࡢ HP1/Swi6 ࡢศᕸࢆㄪࡓࡇࢁࠊᴟࡵ࡚ ⯆ ῝ ࠸ ㌿ ⤊ ⤖ Ⅼ 㸦 TTS: Transcriptional Termination Site㸧㏆ഐ㧗ᗘ⃰⦰ࡋ࡚⤖ྜࡋ࡚࠸ ࡿࡀ᫂ࡽ࡞ࡗࡓࠋࡇࡢ HP1/Swi6 ࡢ TTS ࡢ ⃰⦰ࡣ࣊ࢸࣟࢡ࣐ࣟࢳࣥୖ࠾ࡅࡿ HP1/Swi6 ࡢ⤖ ྜᵝᘧࡣ␗࡞ࡾࠊH3K9me ࡲࡗࡓࡃ౫Ꮡࡋ࡚࠾ ࡽࡎࠊHP1/Swi6 ࡢᣢࡘ DNA ⤖ྜάᛶࡼࡗ࡚ TTS ⃰⦰ࡉࢀ࡚࠸ࡿࡀ᥎ ࡉࢀࡿࠋ FACT ࡢ ChIPѸseq ゎᯒࡣ FACT ᵓᡂᅉᏊࡢ୰ ࡛ࡶࣄࢫࢺࣥ H2A/H2B ࢩࣕ࣌ࣟࣥάᛶࢆᣢࡕࠊ HP1/Swi6 ≀⌮ⓗ⤖ྜࡍࡿ Spt16 ࢆ⏝࠸ࡓࠋ࣊ ࢸࣟࢡ࣐ࣟࢳࣥୖࡢ Spt16 ⤖ྜࡣ swi6+㑇ఏᏊ◚ ቯᰴ࡛㒊ศⓗᦆ࡞ࢃࢀ࡚࠸ࡿࡀ᫂ࡽ࡞ࡾࠊ HP1/Swi6 ౫ Ꮡ ࡋ ࡓ ࣊ ࢸ ࣟ ࢡ ࣟ ࣐ ࢳ ࣥ ୖ ࡢ FACT ࡢࣜࢡ࣮ࣝࢺ࣓ࣥࣥࢺᶵᵓࡀࡅࡉࢀࡓࠋ ࡲࡓ࣮ࣘࢡ࣐ࣟࢳࣥୖ࡛ࡣ HP1/Swi6 ࡀ㧗ᗘ⃰⦰ ࡍࡿ TTS ㏆ഐᑐࡋ࡚ࡸࡣࡾ FACT ࡶ⃰⦰ࡉࢀ࡚࠸ ࡿࡀ᫂ࡽ࡞ࡗࡓࠋࡲࡓࡇࡢࠊswi6+㑇ఏᏊࡢ Ḟኻࡸ HP1/Swi6 ࡢ⤖ྜࡀษࢀࡿ Spt16 ኚ␗యࡢ ศᕸࢆほᐹࡍࡿ TTS ࡢ㧗ᗘ࡞⃰⦰ࡀゎᾘࡉࢀࠊ gene body ᑐࡋ࡚ࡰᆒ୍ศᕸࡍࡿࡼ࠺࡞ኚ ࡀほᐹࡉࢀࡓࠋࡲࡓ RNA-seq ࡛ᚓࡽࢀࡓ㌿⏘≀ࡢ ゎᯒࡽ swi6+㑇ఏᏊḞኻᰴࡸ FACT ኚ␗ᰴ࡛ࡶࢭ ࣥࢫ㙐ࡢ㌿⤊⤖ࡣṇᖖ㉳ࡁࡿࡇࡀほᐹࡉࢀࡓࠋ ࡋࡋࠊࡑࡢ୍᪉࡛ HP1/Swi6 ࡸ FACT ࡀ㧗ᗘ TTS ✚ࡋ࡚࠸ࡓ㑇ఏᏊ⩌࡛ࡣࣥࢳࢭࣥࢫ㙐ࡢ ㌿⏘≀㔞ࡀቑຍࡋ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ㏆ᖺࡢሗ࿌࡛ࡣࣄࢫࢺࣥ H3K9me ౫Ꮡⓗ࣊ࢸࣟ ࢡ࣐ࣟࢳࣥᙧᡂࡣ㠀ᖖከᒱΏࡿᅉᏊ⩌ࡢ㛵 ࡀ♧ࡉࢀ࡚࠸ࡿࠋຍ࠼࡚ࡑࢀࡽࡢᅉᏊ⩌ࡣ࣊ࢸࣟࢡ ࣐ࣟࢳࣥᙧᡂࡢࡳ࡞ࡽࡎࢡ࣐ࣟࢳࣥไᚚ࡞ࡾࢢ ࣮ࣟࣂࣝ㛵ࢃࡗ࡚࠸ࡿࢆ♧၀ࡍࡿ⤖ᯝࡀሗ࿌ࡉ ࢀጞࡵ࡚࠸ࡿࠋ࠼ࡤ࣊ࢸࣟࢡ࣐ࣟࢳࣥࣛࣥࢻ ࡤࢀࡿ H3K9me ࡢᑠつᶍ࡞ศᩓࡀ࣮ࣘࢡ࣐ࣟ ࢳࣥୖ࡛ぢࡽࢀࡿ㇟ࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀࡑࢀࡀᣢ ࡘᮏ㉁ⓗ࡞ពࡣᮍࡔ࡛᫂࠶ࡿࠋᮏᨭ࡛◊✲ ᑐ㇟ࡋࡓ HP1/Swi6 ࡣ H3K9me ౫Ꮡࡋ࡞࠸≧ែ ࡛㑇ఏᏊࡢ TTS ㏆ഐ㧗ᗘ⃰⦰ࡉࢀ࡚࠾ࡾࠊࡑࡢ ᒁᅾࡣ FACT ┦౫Ꮡࡋ࡚࠸ࡿࠋTTS ㏆ഐ㧗ᗘ ✚ࡍࡿࡼ࠺࡞ᅉᏊࡣ⌧ᅾࡲ࡛࠶ࡲࡾࡀ࡞ࡃࠊ TTS ㏆ഐ࡛ఱࡀ㉳ࡁ࡚࠸ࡿࢆヲ⣽ゎᯒࡍࡿⅭࡢ ࡁ࡞㊊ࡾ࡞ࡿᒎᮃ࡛ࡁࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Suzuki S. et al., Nucleic Acids Research, in press - 409 - බເᨭㄢ㢟㸸2010 ᖺᗘࠊ2011 ᖺᗘ ⬚⭢࠾ࡅࡿ⮬ᕫᙧᡂ⮬ᕫㄆ㆑ ≉ᐃ㡿ᇦ◊✲ 㧘ℊ ὒ࣭ᚨᓥᏛᝈࢤࣀ࣒◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 CDR3 㡿ᇦࡢ㛗ࡣㄞࡳษࡿࡇࡀᅔ㞴࡛࠶ࡗࡓࡓ ⋓ᚓච⣔ࡼࡿ⏕య㜵ᚚࡣࠊࣜࣥࣃ⌫ࡢᢠཎ ཷᐜయࡀ᭷⏝࡞ᢠཎㄆ㆑≉␗ᛶࣞࣃࢺࢆ☜❧ࡍࡿ ࡇࡀྍḞ࡛࠶ࡿࠋT ⣽⬊ࡢᢠཎཷᐜయ㸦TCR㸧 ࣞࣃࢺࡣࠊ⬚⭢⓶㉁࡛ࡢṇࡢ㑅ᢥࠊࡑࢀࡘ࡙ ࡃ⬚⭢㧊㉁࡛ࡢ⮬ᕫᐶᐜ☜❧ࢆࡍࡿࠊከẁ㝵ࡢ ⣽⬊㐠ศᒱไᚚࢆ㏻ࡌ࡚ᙧᡂࡉࢀࡿࠋᡃࠎࡣ᭱㏆ࠊ ᭷⏝ CD8T ⣽⬊ࡢṇࡢ㑅ᢥᚲ㡲࡞ࠕ⬚⭢ࣉࣟࢸ ࢯ࣮࣒ࠖࡢᅛ᭷ᵓᡂ㙐 ǃW ࢆḞᦆࡍࡿ࣐࢘ࢫ࡛ࡣ CD8T ⣽⬊ࡢṇࡢ㑅ᢥ␗ᖖࡀ⏕ࡌࠊ୍㒊ࡢࣟ ࡵࠊࢱࢢ㓄ิࡢ┤ᚋ㸲࣑ࣀ㓟㸦CDR3N ᮎ➃㸧 ↔Ⅼࢆ⤠ࡾࠊǃW Ḟᦆ⩌ᑐ↷⩌ࡢ㛫࡛㓄ิࢆẚ㍑ࡋ ࡓࠋ㸰ᅇࡢᐇ㦂ࡽࠊ9ǃ ฟ⌧㢖ᗘ୧⩌ࡢᕪࡀ㢧ⴭ ࡞ 9ǃ ࡸ 9ǃ ࢆྵࡴ」ᩘࡢ CDR3 ࠾࠸࡚㓄ิࡢ ᫂ࡽ࡞㐪࠸ࡀㄆࡵࡽࢀࡓࠋࡲࡓࠊ୍㒊ࡢ㓄ิ㸦9ǃ ࠾ࡅࡿ ASRT ࡸ 9ǃ ࠾ࡅࡿ ASSY㸧ࡣࠊǃW Ḟᦆ࣐࢘ࢫ࡛ࡃ᳨ฟࡉࢀ࡞ࡗࡓࠋࡇࡢࡼ࠺ࠊ ࠕ᭷⏝ࣞࣃࢺࠖࠕᙺ❧ࡓࡎࣞࣃࢺࠖࡢ୍➃ࢆ ᥥฟࡍࡿࡇࡀ࡛ࡁࡓࠋ୍᪉ࠊᚓࡽࢀࡿ TCR 㓄ิࡀ ᛂᛶࡸ࢘ࣝࢫᢠᛶḞኻࡀ࠶ࡿࡇࢆ᫂ࡽ ୍ᗘࡢゎᯒ࡛ 1000 ⛬ᗘᑡᩘࠊ࣮ࣜࢻ㛗ࡀ▷ࡃ ࡋࡓࠋࡋࡋࠊTCR ࣞࣃࢺࡀศᏊᵓ㐀࠾࠸࡚ 㛗 CDR3 㓄ิࢆᚓࡿࡇࡀᅔ㞴ࠊ࠸ࡗࡓᢏ⾡ⓗ࡞ ࡢࡼ࠺ኚᐜࡋ࡚࠸ࡿࢆලయⓗᥥฟࡍࡿࡣ⮳ ࡗ࡚࠸࡞࠸ࠋᮏ◊✲࡛ࡣࠊ ࠕ᭷⏝ࣞࣃࢺࠖࢆࡶࡘṇ ၥ㢟Ⅼࡀุ᫂ࡋࡓࠋᚋࡇࢀࡽࡢၥ㢟ࢆඞ᭹ࡋࡘࡘࠊ TCR ࣞࣃࢺࠕ᭷⏝ᛶࠖࡢᐜゎ᫂ࢆ㐍ࡵࡿࠋ ᖖ࣐࢘ࢫࡢ CD8T ⣽⬊ࠊࣟᛂᛶࡸᢠ࢘ࣝࢫ ᢠᛶࢆ࠺ࡲࡃⓎࡍࡿࡇࡢ࡛ࡁ࡞࠸ࠕᙺ❧ࡓࡎ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣞࣃࢺࠖࢆࡶࡘ ǃW Ḟᦆ࣐࢘ࢫࡢ CD8㹒⣽⬊ࡽ ࡇࢀࡲ࡛ࠊTCR ࣞࣃࢺࡣᴟࡵ࡚ከᵝ࡛࠶ࡿࡢ ᚓࡽࢀࡓ RNA ࢆࡶࠊḟୡ௦ᆺ DNA ࢩ࣮ࢣࣥࢧ ㄆ㆑ࡣඹ᭷ࡉࢀࡘࡘࡶࠊࡑࡢศᏊᵓ㐀ࡢᐇീࡣ᫂ࡽ ࡼࡿ⥙⨶ⓗ㓄ิỴᐃࡼࡗ࡚ TCRǃV 㡿ᇦࡢ N ᮎ ࡛ࡣ࡞ࡗࡓࠋᮏ◊✲ࡣࠊḟୡ௦ᆺࢩ࣮ࢣࣥࢧ ➃ഃ㓄ิࢆᢳฟࡋࠊᢠཎㄆ㆑ከᵝᛶ㒊㓄ิሗࢆ ࡼࡿゎᯒࢆ⏝࠸࡚ࠊ⭾࡞ከᵝᛶࢆ᭷ࡍࡿ TCR ࣞࣃ ࢺࡢゎ᫂ࢆ┠ᣦࡍ᪂ࡓ࡞ヨࡳ࡛࠶ࡗࡓࠋࡇࡢᗘࡢ ⥙⨶ⓗẚ㍑ࡍࡿࡇࢆ┠ⓗࡋࡓࠋ ᨭࢆཷࡅ࡚ࠊࡑࡢᇶᮏⓗᡭἲࢆ☜❧ࡍࡿࡶࠊ 㸺ᨭࡢෆᐜ㸼 ᨵၿࡍࡁㄢ㢟ࢆ᫂ࡽࡍࡿࡇࡀ࡛ࡁࡓࠋᚋࠊ ṇᖖ࣐࢘ࢫࡢ CD8T ⣽⬊ ǃW Ḟᦆ࣐࢘ࢫࡢ CD8T ⣽⬊ࡽᚓࡽࢀࡓ RNA ࢆࡶࠊᮾிᏛࡢ Ⳣ 㔝 ༤ ኈ 㕥 ᮌ ༤ ኈ ࡽ ࡢ ᨭ ࡼ ࡗ ࡚ Illumina Genome Analyzer ࡼࡿ RNA-seq ゎᯒࢆ⾜࡞ࡗࡓࠋ ࡲࡓᚓࡽࢀࡓࢹ࣮ࢱࡽࢥࣥࣆ࣮ࣗࢱゎᯒࢆ⾜࠸ࠊ TCRǃ ᢠཎㄆ㆑ከᵝᛶ㡿ᇦ㸦CDR3 㡿ᇦ㸧ࡢ㓄ิ ሗࢆ⥙⨶ⓗ᳨⣴ࡋࡓࠋ ᮏ◊✲࡛ᚓࡓ⤖ᯝࢆᇶᢏ⾡ࢆᨵⰋࡋࡘࡘゎᯒࢆ㐍 ࡵࡿࡇࡼࡗ࡚ࠊ⏕యࢆ㜵ᚚࡍࡿචࢩࢫࢸ࣒ ྍḞࡢ TCR ࣞࣃࢺࠕ᭷⏝ᛶࠖࡢᐇయศᏊᇶ┙ ࢆ࠼ࡿࡶࠊࡑࡢኚᐜࢆᐈほⓗホ౯ࡍࡿᢏ ⾡ࢆ☜❧ࡋ࡚࠸ࡃ┠㏵ࡀ❧ࡗࡓࠋᑗ᮶ⓗ T ⣽⬊ࣞ ࣃࢺࡢேⅭⓗไᚚࡼࡿච࣏ࢸࣥࢩࣕࣝࡢᙉࠊ ච㛵㐃ᝈࡢ᰿ⓗ⒪ἲࡢ㛤Ⓨࡘ࡞ࡀࡿᮇ ᚅࡉࢀࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᚓࡽࢀࡓ RNA-seq ゎᯒࢹ࣮ࢱࡘ࠸࡚ࠊTCRVǃ 㙐 33 ✀ࡑࢀࡒࢀࡢ N ᮎ➃㒊ศࢆࢱࢢ㓄ิࡋ࡚ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⏝ࡍࡿࡇࡼࡾࠊT&5ǃ 㙐 CDR3 㡿ᇦࡢከᵝᛶࢆ Takada K, et al. Nature Immunology. 16, 1069-1076, ࣑ࣀ㓟㓄ิሗࡼࡾᥥฟࡍࡿࡇᡂຌࡋࡓࠋ 2015. - 410 - බເᨭㄢ㢟㸸2014 ᖺᗘࠊ2015 ᖺᗘ ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫ࠾ࡅࡿศ⠇ RNA ࢤࣀ࣒ࣃࢵࢣ࣮ࢪࣥࢢᶵᵓࡢゎᯒ㸦2014 ᖺᗘ㸧 ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫศ⠇ࢤࣀ࣒ࡢ⢏Ꮚෆ❧య㓄⨨Ỵᐃ㸦2015 ᖺᗘ㸧 ⱝᡭ◊✲㸦㹀㸧 ἑ ┤ᕫ࣭ᚤ⏕≀Ꮫ◊✲ᡤ➨ 3 ⏕≀άᛶ◊✲㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫࡣ࣐ࢼࢫ㙐 51$ ࢆࢤࣀ ࣒ࡋ࡚ᣢࡕࠊࢤࣀ࣒ࡣ ᮏศ⠇ࡉࢀ࡚࠸ࡿࠋ ✀ ᮏࡢ࢘ࣝࢫࢤࣀ࣒ 51$ ࡀ୍ࡘࡢ⢏Ꮚࣃࢵࢣ ࣮ࢪࣥࢢࡉࢀࡿᶵᵓࡣศ⠇ࡢ⤌࠼࡛ฟ⌧ࡍࡿࣃࣥ ࢹ࣑ࢵࢡࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫไᅽ࠸࠺Ⅼ࡛㔜 せ࡞ࡿ⪃࠼ࡽࢀࡿࡀࠊࡑࡢศᏊᶵᵓࡣ࡛᫂࠶ ࡿࠋ⩻ヂ㡿ᇦࡶྵࡵࡓ࢘ࣝࢫࢤࣀ࣒ୖࡢ୍㒊㓄ิ ࡀຠ⋡ⓗ࡞ࣃࢵࢣ࣮ࢪࣥࢢᚲせ࡛࠶ࡿࡇࡽࠊ ศ⠇㛫ࡢㄆ㆑ࡣ 51$51$ ┦స⏝ࡀᐃࡉࢀ࡚࠸ ࡿࡀࠊࡑࡢ᭷↓ࡘ࠸࡚ド᫂ࡣ࡞ࡉࢀ࡚࠸࡞࠸ࠋࡑ ࡇ࡛ᮏ◊✲࡛ࡣࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫศ⠇ࢤࣀ ࣒ࡢ㑅ูࠊࣃࢵࢣ࣮ࢪࣥࢢᶵᵓࢆ᫂ࡽࡍࡿࡇ ࢆ┠ⓗࡋ࡚ࠊࣃࢵࢣ࣮ࢪࣥࢢᚲせ࡞⩻ヂ㡿ᇦ ෆࡢሷᇶࠊศ⠇㛫 51$51$ ┦స⏝ࡢ᭷↓ࡶྵࡵ ࡓ࢘ࣝࢫ⢏Ꮚෆࢤࣀ࣒ศ⠇ࡢ❧య㓄⨨ࢆ᫂ࡽ ࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࣃࢵࢣ࣮ࢪࣥࢢᚲせ࡞⩻ヂ㡿ᇦෆࡢሷᇶࡢྠᐃ ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫࢱࣥࣃࢡ㉁࡛࠶ࡿ +$ ࡲࡓ ࡣ 0 ࢆࢥ࣮ࢻࡍࡿศ⠇⤊Ṇࢥࢻࣥࢆᤄධࡋࠊ⤊Ṇ ࢥࢻࣥࡢୗὶഃ㓄ิࡢ୍㒊ࢆࣛࣥࢲ࣒࡞㓄ิ⨨ࡁ ࠼ࡓ⤌࠼࢘ࣝࢫࣉ࣮ࣝࢆసᡂࡋࡓࠋసᡂࡋࡓ ࢘ࣝࢫࣉ࣮ࣝࢆ 0'&.+$ ࡲࡓࡣ 0'&.0 ⣽⬊࡛⥅ ௦ࡋࠊ⥅௦ ௦┠ࠊ ௦┠ࠊ ௦┠ࡢ࢘ࣝࢫࡽ࢘ ࣝࢫ 51$ ࡢ⢭〇ࢆ⾜࠸ࠊᤄධࡋࡓࣛࣥࢲ࣒㓄ิ㡿 ᇦࢆྵࡴ㡿ᇦࢆ 573&5 ࡼࡾቑᖜࡋࠊ0L6HT ࡼࡾ $PSOLFRQVHT ࢆ⾜࠸ࠊ⥅௦ࡼࡾ㑅ᢥࡉࢀࡓ㓄ิ ࡘ࠸࡚ゎᯒࢆ⾜ࡗࡓ㸦 ᖺᗘ㸧ࠋ ࢘ࣝࢫ⢏Ꮚෆࢤࣀ࣒ศ⠇ࡢ❧య㓄⨨ࡢゎ᫂ ࢘ࣝࢫ⢏Ꮚෆ࡛ࡢศ⠇㓄⨨ࢆỴᐃࡍࡿࡓࡵ」 ᩘ㑇ఏᏊࡢ㏆᥋㡿ᇦࢆ⥙⨶ⓗ᫂ࡽࡍࡿ &VHT ࢵࢭࢆཧ⪃ࡋ࡚ࠊ⢭〇ࣥࣇ࢚ࣝࣥࢨ࢘ࣝ ࢫ⢏Ꮚࡢࢡࣟࢫࣜࣥࢡ࢘ࣝࢫ㏱㐣ฎ⌮51$ 㒊ศ ษ᩿51$OLJDWLRQ ࡢ⣔ࢆᵓ⠏ࡋࡓࠋࣛࣈ࣮ࣛࣜస 〇ᚋࠊ0L6HT ࡼࡿつᶍࢩ࣮ࢡ࢚ࣥࢩࣥࢢࢆ⾜ࡗࡓࠋ ㄞ ࡳྲྀࡗ ࡓࣜ ࣮ࢻࡢ ゎᯒࢆ ⾜࠸ࠊ ศ⠇ 51$ 㛫࡛ OLJDWLRQ ࡀ⾜ࢃࢀ࡚࠸ࡿ࣮ࣜࢻࢆᢳฟࡋࠊ⢏Ꮚෆ࡛ ㏆㊥㞳࠶ࡿศ⠇㛫㡿ᇦࡢྠᐃࢆ⾜ࡗࡓࠋ⣔ࡢᵓ⠏ ࢆ☜ㄆᚋࠊ+L6HT ࡼࡾ᭦つᶍࢹ࣮ࢱࡢ㞟ࢆ⾜ ࡗࡓ㸦ࠊ ᖺᗘ㸧 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㑅ᢥࢆཷࡅࡿࣛࣥࢲ࣒㓄ิࡘ࠸࡚ゎᯒࢆ⾜ࡗࡓ ⤖ᯝࠊHA ࢆࢥ࣮ࢻࡍࡿศ⠇ࡘ࠸࡚ࡣ≉ᐃࡢሷᇶࡀ ᙉࡃ㑅ᢥࢆཷࡅࡿࡇࡣ࡞ࡃࠊࣛࣥࢲ࣒㓄ิ㒊 ྵࡲࢀࡿሷᇶẚ⋡ࡣ⥅௦ 1 ௦┠ࡽ⥅௦ 5 ௦┠ࡲ࡛ ࡰྠ⛬ᗘࡢྜ࡛࠶ࡗࡓࠋM2 ࢆࢥ࣮ࢻࡍࡿศ⠇ ࡘ࠸࡚ࡣ࠸ࡃࡘࡢሷᇶࡀ⥅௦ 5 ௦┠ࡲ࡛≉ᐃࡢ ሷᇶ㑅ᢥࡉࢀ࡚࠸ࡓࠋ≉ᐃࡢሷᇶ㑅ᢥࢆཷࡅࡓ 㒊ኚ␗ࢆᣢࡘ⤌࠼࢘ࣝࢫࢆసᡂࡋࠊ࢘ࣝ ࢫቑṪࡘ࠸࡚㔝⏕ᆺ࢘ࣝࢫẚ㍑ࡋࡓ⤖ᯝࠊ⤌ ࠼࢘ࣝࢫࡣ㔝⏕ᆺ࢘ࣝࢫẚ㍑ࡋ࡚ឤᰁᛶ࢘ ࣝࢫ⏘⏕㔞ࡀῶᑡࡋ࡚࠸ࡓࠋࡇࢀࡽ⤖ᯝࡣࠊつ ᶍࢩ࣮ࢡ࢚ࣥࢫࡼࡾỴᐃࡋࡓ㑅ᢥࢆཷࡅࡿሷᇶࡀ ࢘ࣝࢫቑṪ㛵ࡍࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋ ⢭〇ࣥࣇ࢚ࣝࣥࢨ࢘ࣝࢫᑐࡋ࡚ࢡࣟࢫࣜࣥ ࢡ-࢘ࣝࢫ㏱㐣ฎ⌮-RNA 㒊ศษ᩿-RNA ligation ᚋࣛࣈ࣮ࣛࣜࡢㄪᩚࢆ⾜࠸ࠊMiSeq ࡼࡾㄞࡳྲྀ ࡾࢆ⾜࠺ࡇ࡛ࠊࢤࣀ࣒ RNA ศ⠇㛫࡛ࣛࢤ࣮ࢩࣙ ࣥࡀ㉳ࡇࡗࡓ㒊ࡘ࠸࡚ゎᯒࢆ⾜ࡗࡓࠋMiSeq ࡼࡿㄞࡳྲྀࡾࡢ⤖ᯝࠊᑡ࡞࠸࣮ࣜࢻ࡛ࡣ࠶ࡿࡀࠊศ ⠇㛫࡛ࣛࢤ࣮ࢩࣙࣥࡀ㉳ࡇࡗࡓ࣮ࣜࢻࢆ☜ㄆࡋࠊ ࣐ࢵࣆࣥࢢࢆ⾜ࡗࡓࠋ࣐ࢵࣆࣥࢢࡢ⤖ᯝࠊࢤࣀ࣒ศ ⠇ RNA ࡢᮎ➃㡿ᇦ-ᮎ➃㡿ᇦࡢࡳ࡛࡞ࡃࠊศ⠇ RNA ࡢ୰ኸ㡿ᇦ-ᮎ➃㡿ᇦ࡛ࣛࢤ࣮ࢩࣙࣥࡀ㉳ࡇࡿࣜ ࣮ࢻࡀ☜ㄆࡉࢀࡓࠋࡇࢀࡽ⤖ᯝࡼࡾࠊࣥࣇ࢚ࣝࣥ ࢨ࢘ࣝࢫ⢏Ꮚෆ࡛ࢤࣀ࣒ RNA ศ⠇ࡣ୍᪉ྥ࡛ࡣ ࡞ࡃ࡚ࠊ㏫᪉ྥ࡛ࡶࣃࢵࢣ࣮ࢪࣥࢢࡉࢀࡿࡀ᫂ࡽ ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᅇࡢ⤖ᯝࡼࡾࠊ࢘ࣝࢫࢱࣥࣃࢡ㉁ࡢኚ␗ ࡣ↓㛵ಀ࡞ RNA ࡢኚ␗ࡀ࢘ࣝࢫቑṪᙳ㡪ࢆ ࠼ࡿࡇࢆ᫂ࡽࡋࡓࠋᚋࠊᚓࡽࢀࡓ㓄ิᑐ ࡋ࡚᭦ゎᯒࢆ⾜࠺ࡇ࡛ࠊ࢘ࣝࢫ⢏Ꮚ୰ࣃࢵ ࢣ࣮ࢪࣥࢢࡉࢀࡿ㓄ิࡘ࠸࡚ࡼࡾヲ⣽᫂ࡽ ࡍࡿணᐃ࡛࠶ࡿࠋࡲࡓࠊᅇྠᐃࡋࡓሷᇶࡀ࢘ࣝ ࢫቑṪ࠾࠸࡚ᯝࡓࡍᙺࡘ࠸࡚᫂ࡽࡍࡿண ᐃ࡛࠶ࡿࠋ ࢘ࣝࢫ⢏Ꮚෆࢤࣀ࣒ศ⠇ࡢ❧య㓄⨨ゎ᫂ࡘ࠸ ࡚ࡣࠊࢡࣟࢫࣜࣥࢡ-࢘ࣝࢫ㏱㐣ฎ⌮-RNA 㒊ศษ ᩿-RNA ligation ࡢ⣔ࡣ☜❧ࡋ࡚࠸ࡿࡢ࡛ࠊHiSeq ࡼࡿࢩ࣮ࢡ࢚ࣥࢫࢆ⾜࠸࣮ࣜࢻᩘࢆቑࡸࡍࡇ࡛ࡼ ࡾヲ⣽ศ⠇㛫 RNA ࡢ㏆᥋㒊ࢆ᫂ࡽࡍࡿண ᐃ࡛࠶ࡿࠋᚓࡽࢀࡓ⤖ᯝࢆヲ⣽ゎᯒࡍࡿࡇ࡛ࠊ ሷᇶࣞ࣋ࣝ㏆࠸ᙧ࡛ศ⠇㛫┦స⏝㒊ࢆྠᐃ࡛ ࡁࡿ⪃࠼࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 411 - බເᨭㄢ㢟㸸2012㹼2013 ᖺᗘ 㨶㢮ࡢᾏỈ㐺ᛂࢆᨭ㓄ࡍࡿ⬻⭠ࡢෆศἪ┦㛵 ᇶ┙◊✲㸦㸿㸧 ➉ ⚈㑻࣭ᮾிᏛẼᾏὒ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ᾏỈ㨶ࡣࠊ⬺Ỉᑐฎࡍࡿࡓࡵ┒ࢇᾏỈࢆ㣧ࡳࠊ 㣧ࢇࡔᾏỈࡢ⣙ 90㸣ࢆ⭠࡛྾ࡋ࡚࠸ࡿࠋࡲࡓࠊయ ෆධࡍࡿ NaCl ࢆ㪰ࡢሷ㢮⣽⬊࡛⃰⦰ࡋ࡚ฟ ࡋࠊ㧗ሷศ⎔ቃ࡛యᾮࣂࣛࣥࢫࢆಖࡗ࡚࠸ࡿࠋࡇࡢ ࡼ࠺ࠊ⭠㪰ࡣ㨶㢮ࡢ᭱ࡶ㔜せ࡞ᾐ㏱ᅽㄪ⠇ჾᐁ ࡛࠶ࡿࡀࠊ⭠ࡸ㪰࠾ࡅࡿᾏỈ㐺ᛂࢆᨭ㓄ࡍࡿ㑇ఏ Ꮚࡣࡲࡔぢࡘࡗ࡚࠸࡞࠸ࠋᮏ◊✲࡛ࡣࠊῐỈᾏ Ỉ᪉㐺ᛂࡍࡿࡇࡀ࡛ࡁࠊࢤࣀ࣒ሗࡀබ⾲ࡉ ࢀ࡚࠸ࡿ࢘ࢼࢠ࣓ࢲ࢝ࢆ⏝࠸࡚ࠊࡇࢀࡽࡢჾᐁ࡛ Ⓨ⌧ࡋࠊᾏỈ⛣⾜ࡼࡾⓎ⌧ࡀ᭦᪂ࡍࡿ㑇ఏᏊࢆྠ ᐃࡍࡿࡇࢆ┠ⓗࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᾏỈ㐺ᛂ㑇ఏᏊࢆྠᐃࡍࡿࡓࡵࠊ࢘ࢼࢠ࣓ࢲ࢝ࢆ ῐỈࡽᾏỈ⛣⾜ࡉࡏࠊ⛣⾜ᚋ 1, 3 㛫㸦ึᮇ㐺 ᛂ㑇ఏᏊ㸧ࠊ࠾ࡼࡧ 1, 7 ᪥㸦㛗ᮇ㐺ᛂ㑇ఏᏊ㸧⭠ 㪰ࢆྛ 5 ༉ࡎࡘࢧࣥࣉࣜࣥࢢࡋ࡚ࠊPolyA RNA ᢳฟ ࢆ⾜ࡗࡓࠋⓎ⌧㑇ఏᏊゎᯒࡣ㕥ᮌࢢ࣮ࣝࣉࡢ㠃ⓗ ࡞ᨭࢆཷࡅ㸦 120 ࢧࣥࣉࣝ㸧 ࠊRNA-seq ゎᯒࡣ ᒾᓮࢢ࣮ࣝࣉࡼࡾ⤯࡞ࡿᨭࢆཷࡅࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᾏỈῐỈ᪉㐺ᛂ࡛ࡁࡿᗈሷᛶ㨶࡛࠶ࡿࡀࠊ㉳ ※ⓗᾏỈ㨶࡛࠶ࡿ࢘ࢼࢠῐỈ㨶࡛࠶ࡿ࣓ࢲ࡛࢝ ࡣࠊᾏỈ⛣⾜ࡼࡾ␗࡞ࡿᛂࢆ♧ࡍ㑇ఏᏊࡀከᩘ ぢࡘࡗࡓࠋࡲࡓࠊᾏỈ┤᥋ࡉࡽࡉࢀࡿ㪰ࠊᾘ ⟶๓㒊ࡼࡿ⬺ሷ࡞࡛⤌ᡂࡀኚࡋࡓᾏỈࢆཷ ᐜࡍࡿ⭠࡛ࡣࠊᾏỈ⛣⾜ᛂࡍࡿ㑇ఏᏊ᫂ࡽ ࡞㐪࠸ࡀ࠶ࡗࡓࠋࢤࣀ࣒ሗࡀഛࡋ࡚࠸ࡿ࣓ࢲ࢝ ࡢ⭠ࡢึᮇᛂ㑇ఏᏊࡣࢫࢺࣞࢫ㑇ఏᏊࡀከᩘ࠶ ࡾࠊῐỈࡽῐỈ⛣⾜ࡋࡓᑐ↷⩌࡛ࡶୖ᪼ࡋࡓࠋ ࡋࡋࠊᾐ㏱ᅽࡢୖ᪼ࡢࡳ≉␗ⓗᛂࡍࡿ㑇ఏ Ꮚࢆ 5 ✀ᚓࡿࡇࡀ࡛ࡁࡓࠋᾏỈ⛣⾜ᐇ㦂ᛂࡋ ࡓࡍ࡚ࡢ㑇ఏᏊ㛫ࡢ┦㛵ಀࢆ String Analyses ࡛ゎᯒࡋࡓࡇࢁࠊ࠸ᐦ᥋┦㛵ࡋ࡚࠸ࡿࡇ ࡀ᫂ࡽ࡞ࡗࡓࠋࡲࡓࠊᾐ㏱ᅽ≉␗ⓗᛂࡍ ࡿ㑇ఏᏊࡢⓎぢࡣࠊࡇࢀࡲ࡛Ⓨ⾲ࡉࢀࡓ㑇ఏᏊ㸦ࢫ ࢺࣞࢫ㑇ఏᏊ㸧ࡣ␗࡞ࡿࡓࡵࠊ᭱ึࡢㄽᩥࡋ࡚ Ⓨ⾲ࡋࡓࠋ⌧ᅾࠊࡇࢀࡽ㑇ఏᏊࢆ࣓ࢲ࢝ࢆ⏝࠸࡚⭠ ≉␗ⓗࣀࢵࢡࢲ࢘ࣥࡋ࡚ࠊ࣓ࢲ࢝ࡢඃࢀࡓᾏỈ㐺 ᛂ⬟࠾ࡅࡿᙺࢆㄪ࡚࠸ࡿࠋ ᚋࡣࠊᮏᨭ࡛ᚓࡽࢀࡓ㈗㔜࡞ࢹ࣮ࢱࢆ⏝࠸࡚ࠊ 2 ࡘࡢ␗࡞ࡿどⅬࡽᡂᯝࢆࡲࡵ࡚࠸ࡃணᐃ࡛࠶ ࡿࠋ➨ 1 ࡢどⅬࡣࠊ࣓ࢲ࢝࢘ࢼࢠࡢ㐪࠸࡛࠶ࡿࠋ ⭠࠾ࡅࡿึᮇᛂ㑇ఏᏊࡔࡅࢆࡳ࡚ࡶࠊ୧✀࠾ ࠸࡚ඹ㏻ࡢ㑇ఏᏊࡸ␗࡞ࡿ㑇ఏᏊࡀከᩘぢࡘࡗ࡚ ࠸ࡿࠋඹ㏻㑇ఏᏊࡣᾏỈ㐺ᛂ㔜せ࡞㑇ఏᏊ࡛࠶ࡿ ⪃࠼ࡽࢀࡿࡀࠊ␗࡞ࡿ㑇ఏᏊ㛵ࡋ࡚ࡶࠊ㐍Ꮫ ⓗ࡞ 2 ✀ࡢ㐪࠸㛵ࡍࡿ௬ㄝࢆ❧࡚࡚ᐇドᐇ㦂ࢆ⾜ ࡗ࡚࠸ࡿࠋࡲࡓࠊ㍺㏦యࡸ⣽⬊᥋╔ᅉᏊ࡞⤌⧊ࢆ సࡾ᭰࠼ࡿ㛗ᮇᾏỈ㐺ᛂ㑇ఏᏊࡶከࡃࡢ␗࡞ࡿ㑇 ఏᏊࡀぢࡘࡗ࡚࠸ࡿࠋࡑࡢ࠺ࡕࡢ࠸ࡃࡘࡣࡇࢀ ࡲ࡛ᾏỈ㐺ᛂࡢ㛵ࡀࡃ▱ࡽࢀ࡚࠸࡞ࡗࡓ 㑇ఏᏊ࡞ࡢ࡛ࠊ࣓ࢲ࢝ẚ࢘ࢼࢠࡢࡼࡾඃࢀࡓ㐺 ᛂ⬟㛵ࡋ࡚࠸ࡿࡢࢆ᫂ࡽࡋࡓ࠸ࠋ ➨ 2 ࡢどⅬࡣࠊ⭠㪰ࡢ㐪࠸࡛࠶ࡿࠋእ⏺ࡢኚ ┤᥋ᛂࡍࡿ㪰⟶⭍ෆᾮᛂࡍࡿ⭠࡛ࡣึᮇ ᛂ㑇ఏᏊࡁ࡞㐪࠸ࡀ࠶ࡿࡇࡀࢃࡗࡓࠋࡲ ࡓࠊ⭠㪰࡛Ⓨ⌧ࡋ࡚࠸ࡿ㍺㏦యࡸࢳࣕࢿࣝࡣ␗࡞ ࡿࡀࠊྠࡌ㍺㏦య࡛ࡶ␗࡞ࡿࣃࣛࣟࢢࡀⓎ⌧ࡋ࡚࠸ ࡿࡇࡀࢃࡗ࡚ࡁࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡀྛ⤌⧊ࡢ ࡢ⣽⬊Ⓨ⌧ࡋ࡚࠸ࡿࢆ in situ hybridization ࡛ㄪࠊୖ⓶⣽⬊Ⓨ⌧ࡋ࡚࠸ࡿሙྜࡣࡑࢀࡽࢆ ⤌⧊≉␗ⓗࣀࢵࢡࢲ࢘ࣥࡋ࡚ࠊᾏỈ㐺ᛂ⬟ࡢ㛵 ࢆㄪࡘࡘ࠶ࡿࠋࡇࢀࡽࡢᡂᯝࡣࠊᚋ 2-3 ᖺࡢ ࠺ࡕ࠸ࡃࡘࡢ㔜せ࡞ㄽᩥࡋ࡚Ⓨ⾲ࡍࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡇ ࢀ ࡲ ࡛ 㨶 㢮 ࠾ ࠸ ࡚ ࠊ DNA microarray ࡸ Subtractive Differential Display ࡞ࡢ᪉ἲࢆ⏝࠸ ࡚ࠊᾏỈ⛣⾜ᚋⓎ⌧ࡀୖ᪼ࡍࡿึᮇᛂ㑇ఏᏊࡀ ᥈ ⣴ ࡉ ࢀ ࡚ ࡁ ࡓ ࠋ ࡑ ࢀ ࡽ ࡢ ◊ ✲ ࡼ ࡾ Osmotic Transcription Factor 1 (Ostf-1)ࡀࢸࣛࣆࡢ㪰࡛ Ⓨぢࡉࢀࡓࡀࠊࣀࢵࢡࢲ࢘ࣥࢆࡋ࡚ࡶᾏỈ㐺ᛂ⬟ ኚࡀぢࡽࢀ࡞࠸࡞ࠊࡑࡢ㔜せᛶ㛵ࡋ࡚ከࡃࡢ ㆟ㄽࡀ࠶ࡗࡓࠋᅇ 2 ࡘࡢ㨶✀࡛ࡇࡢ㑇ఏᏊࢆㄪ ࡓࡇࢁࠊᾏỈ⛣⾜⩌ࡔࡅ࡛ࡣ࡞ࡃࠊᑐ↷⩌࡛ࡶ Ⓨ⌧ࡢୖ᪼ࡀぢࡽࢀࡓࠋࡍ࡞ࢃࡕࠊࡇࡢ㑇ఏᏊࡣࢫ ࢺࣞࢫ⯡ᛂࡋ࡚Ⓨ⌧ࡀୖ᪼ࡍࡿࡇࡀࢃࡗ ࡓࠋࡇࡢࡼ࠺ࠊ┿ࡢᾏỈ㐺ᛂ㛵ࢃࡿ㔜せ࡞㑇ఏ Ꮚࢆྠᐃࡍࡿࡓࡵࡣࠊ⢭ᗘࡢ㧗࠸㑇ఏᏊゎᯒᐇ㦂 ࢆ⾜࠺ᚲせࡀ࠶ࡿࠋࡲࡓࠊ㨶㢮ࡣ့ங㢮ẚ㍑ࡍࡿ ⏕⌮ᶵ⬟ࡶከᵝᛶࡀࡁ࠸ࡓࡵࠊ1 ✀ࡽᚓࡽࢀ ࡓᐇࡀࡢ㨶✀ࡶᙜ࡚ࡣࡲࡿࡣ㝈ࡽ࡞࠸ࠋ ᅇࡢᨭࡼࡾࠊ࣓ࢲ࢝࢘ࢼࢠࡢ⭠㪰࠾ࡅࡿ ᾏỈ⛣⾜ᛂࡍࡿ⭾࡞㑇ఏᏊࡀᚓࡽࢀࠊᛂࡢ 㛫⤒㐣ࡶ᫂ࡽࡍࡿࡇࡀ࡛ࡁࡓࠋᚋࡣࡇࢀ ࡽ⭾࡞ࢹ࣮ࢱࡽࠊ࡚ࡢ㨶✀ඹ㏻࡞ᾏỈ㐺ᛂ 㑇ఏᏊ㛵ࢃࡿಙ㢗࡛ࡁࡿࢹ࣮ࢱࢆᢳฟࡋ࡚ࠊࡇࡢ ศ㔝ࡢⓎᒎᐤ࡛ࡁࡿಙࡌ࡚࠸ࡿࠋ࠼ࡤࠊᾐ ㏱ ᅽ ≉ ␗ ⓗ ᛂ ࡍ ࡿ ึ ᮇ ᛂ 㑇 ఏ Ꮚ (early response gene)ࢆぢࡘࡅࠊࡑࢀࡀ㌿ㄪ⠇ᅉᏊࡢሙྜ ࡣᶆⓗ㑇ఏᏊࢆ᫂ࡽࡋ࡚ࠊࣉ࣮ࣟࣔࢱ࣮㡿ᇦ ࠶ࡿ㌿ᅉᏊࡀ⤖ྜࡍࡿ㓄ิࢆỴᐃࡍࡿࠋࡑࢀࡀ᫂ ࡽ࡞ࡗࡓሙྜࡣࠊࡑࡢ㓄ิࢆᣢࡘ㑇ఏᏊࢆ᳨⣴ ࡋ࡚ᶆⓗ㑇ఏᏊࢆ᥈⣴ࡋࠊࡇࢀࡲ࡛ࡣ␗࡞ࡿࣉ ࣮ࣟࢳࡢᾐ㏱ᅽㄪ⠇◊✲ࢆᥦࡋࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Wong, M., et al., BMC Genomics 15:1134 (2014) Ando, M., et al., Am. J. Physiol. 307:R653-R663 (2014) - 412 - බເᨭㄢ㢟㸸2012 ᖺᗘ 㹐㹌㸿ศゎ㓝⣲㹘㹡㸱㹦㸯㸰㹟ࡼࡿචไᚚᶵᵓࡢゎᯒ ⱝᡭ◊✲㸦㸿㸧 ➉ෆ ⌮࣭ி㒔Ꮫ࢘ࣝࢫ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ⅖ᛂ⟅ࡣࠊච⣽⬊࠾ࡅࡿࢧࢺ࢝ࣥࢆጞࡵ ࡋࡓ㑇ఏᏊⓎ⌧ࡼࡾㄪ⠇ࡉࢀ࡚࠸ࡿࠋච⣽⬊ ࠾ࡅࡿ㑇ఏᏊㄪ⠇ࡋ࡚ࠊ㌿ไᚚຍ࠼ࠊP51$ Ᏻᐃᛶไᚚࡀ㔜せ࡛࠶ࡿࠋᡃࠎࡣࠊ᪂つ 51$ ศゎ㓝 ⣲࡛࠶ࡿ 5HJQDVH㸦=FKD㸧ࢆྠᐃࡋࠊࡇࡢศᏊ ࡀࠊ㸯㸧࣐࢘ࢫ࠾࠸࡚⮬ᕫචᛶ⅖ᛶᝈࡢⓎ ᢚไ㔜せ࡛࠶ࡿࡇࠊ㸰㸧࣐ࢡࣟࣇ࣮ࢪ࡞ ⮬↛ච⣽⬊ࠊ7 ⣽⬊ࢆጞࡵࡋࡓ⋓ᚓච⣔ࡢάᛶ ᢚไ㔜せ࡛࠶ࡿࡇࠊ㸱㸧5HJQDVH Ḟᦆ࣐ࢡࣟ ࣇ࣮ࢪ࠾࠸࡚ࣥࢱ࣮ࣟ࢟ࣥ㸦,/㸧 ࢆጞࡵ ࡋࡓ P51$ ࡸ⺮ⓑࡢⓎ⌧ࡀቑຍࡋ࡚࠸ࡿࡇࠊ㸲㸧 5HJQDVH ࡣ ,/ ࡞ࡢ P51$ ࢆࡑࡢ ̓㠀⩻ヂ㡿ᇦ ࢆࡋ࡚ᢚไࡋ࡚࠸ࡿࡇࢆ♧ࡋ࡚ࡁࡓࠋࡋࡋ࡞ ࡀࡽࠊ,/ 5HJQDVH 㔜Ḟᦆ࣐࢘ࢫ࠾࠸࡚ࡶ⅖ ᛶᝈࡢⴭ᫂࡞ᨵၿࢆㄆࡵࡎࠊᝈⓎ㛵ࢃࡿ ࣓࢝ࢽࢬ࣒ࡣ࡛᫂࠶ࡗࡓࠋᮏ◊✲࡛ࡣࠊ5HQJDVH ࡢ⋓ᚓච⣔࠾ࡅࡿᙺࡢゎᯒࡽࠊචᛂ⟅ࡢ 51$ ไᚚࡼࡿㄪ⠇࣓࢝ࢽࢬ࣒ࢆ᫂ࡽࡍࡿࢆ ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᮏ◊✲࡛ࡣࠊ5HJQDVH ࡢ⋓ᚓච⣽⬊ࠊ≉ 7 ⣽⬊ ࠾ࡅࡿᙺࢆゎᯒࡍࡿࡓࡵ &UHOR[3 ࢩࢫࢸ࣒ࢆ ⏝࠸ࠊࡲࡎࠊ5HJQDVHIOR[ ࣐࢘ࢫࢆస〇ࡋࡓࠋࡇ ࡢ࣐࢘ࢫࢆࠊ7 ⣽⬊≉␗ⓗ &UH ࢆⓎ⌧ࡍࡿ &'&UH ࡶࡋࡃࡣ /FN&UH ࣐࢘ࢫ㓄ࡋࠊ7 ⣽⬊≉␗ⓗ 5HJQDVH Ḟᦆ࣐࢘ࢫࢆస〇ࡋࡓࠋࡇࡢ࣐࢘ࢫࡣ㌟ ࡛ 5HJQDVH ࢆḞᦆࡍࡿ࣐࢘ࢫྠᵝ㔜⠜࡞⮴Ṛ ᛶ⮬ᕫචᝈࢆⓎࡋࠊ7 ⣽⬊Ⓨ⌧ࡍࡿ 5HJQDVH ࡢ 㔜 せ ᛶ ࡀ ᫂ ࡽ ࡞ ࡗ ࡓ ࠋ ࡲ ࡓ ࠊ 5HJQDVH Ḟᦆ 7 ⣽⬊ࡣ 7 ⣽⬊ཷᐜయ㸦7&5㸧่⃭ ᑐࡍࡿቑṪࡸࢧࢺ࢝ࣥ⏘⏕࡞ࡢᛂࡀஹ㐍ࡋ ࡚࠸ࡓࠋ5HJQDVH ࢱࣥࣃࢡ㉁ࡣ 7&5 ่⃭ᑐࡋࠊ 0$/7 ࣉࣟࢸ࣮ࢮࡼࡾษ᩿ࢆཷࡅ࡚ศゎࡉࢀࠊ 5HJQDVH ࡼࡿ 7 ⣽⬊άᛶᢚไᶵ⬟ࡀࠊචᛂ⟅ ࡢ㐣⛬࡛ࢲࢼ࣑ࢵࢡㄪ⠇ࡉࢀ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋ 5HJQDVH ࡀ 7 ⣽⬊࠾࠸࡚ࡢࡼ࠺࡞ P51$ ࢆᶆⓗ ࡋ࡚ࠊ7 ⣽⬊άᛶࢆㄪ⠇ࡋ࡚࠸ࡿࢆ᫂ࡽࡍ ࡿ┠ⓗ࡛ࠊ㔝⏕ᆺཬࡧ 5HJQDVH Ḟᦆ &'7 ⣽⬊ࢆ ࣐࢘ࢫࡼࡾ༢㞳ࡋࠊP51$ ࢆㄪ〇ࠊ7UDQVFULSWRPH ゎ ᯒࡢᨭࢆ౫㢗ࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㔝⏕ᆺࠊཬࡧ 5HJQDVH Ḟᦆ࣐࢘ࢫ⭁⮚ &'7 ⣽⬊ ࢆ⏝࠸ࡓ 7UDQVFULSWRPH ゎᯒࡢ⤖ᯝࠊ୍⩌ࡢ P51$ ࡀ 5HJQDVH Ḟᦆ 7 ⣽⬊࠾࠸࡚㔝⏕ᆺẚ㍑ࡋ࡚Ⓨ ⌧ୖ᪼ࡋ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࡢ P51$ ࡢ ୰ࡣࠊ࣐ࢡࣟࣇ࣮ࢪ࡛ࡇࢀࡲ࡛ྠᐃࡉࢀ࡚࠸ ࡓ ,O ࡞ຍ࠼ࠊF5HO ࡸࠊ,FRVࠊ2[ࠊ,O ࡞ ࡢ P51$ ࢆぢࡘࡅࡿࡇࡀฟ᮶ࡓࠋ᭦ࠊ&'7 ⣽⬊ ࢆࢼ࣮ࣈࠊ࢚ࣇ࢙ࢡࢱ࣮ࡢ ✀㢮ศࡅ࡚ 7UDQVFUSWRPH ゎᯒࢆࡋࡓࡁࡶྠᵝࡢ⤖ᯝࢆᚓࡓࠋ ࡇࡇ࡛᫂ࡽ࡞ࡗࡓ㑇ఏᏊࢆ࣏࣮ࣞࢱ࣮ࢩࢫࢸ࣒ ࡞ࡼࡾ᳨ウࢆຍ࠼ࠊࡇࢀࡽࡢศᏊࢆࢥ࣮ࢻࡍࡿ P51$ ࡀ 5HJQDVH ࡼࡿ┤᥋ࡢᶆⓗ࡞ࡗ࡚࠸ࡿࡇ ࢆ♧ࡍࡀฟ᮶ࡓࠋࡇࡢࡼ࠺ࠊ7UDQVFULSWRPH ゎᯒࡼࡾࠊ⣽⬊✀≉␗ⓗ࡞ 5HJQDVH ᶆⓗ P51$ ࢆ ⥙⨶ⓗ᳨ウࡋࠊྠᐃࡍࡿࡇࡀฟ᮶ࡓࠋ ࡲࡓࠊᖺࡣࠊ5HJQDVH ࡢᶆⓗ P51$ ⤖ྜ≉␗ᛶࢆ ᫂ࡽࡍࡿࡓࡵࠊ+H/D ⣽⬊ )ODJ ࡛ࢱࢢࢆࡋࡓ 5HJQDVH ࢆᏳᐃⓗⓎ⌧ࡉࡏࡓ⣽⬊ᰴࢆస〇ࡋࠊ )ODJ ࡛චỿ㝆ࠊ7UDQVFULSWRPH ゎᯒ㸦5,36HT ゎᯒ㸧 ࢆ⾜࠸ࠊ୍⩌ࡢ 5HJQDVH ᶆⓗ P51$ ࢆᚓࡿࡇࡀฟ ᮶ࡓࠋࡇࢀࡣྠᵝᨭࢆ㡬࠸ࡓ 5RTXLQ ᶆⓗ P51$ ᭷ព࣮࢜ࣂ࣮ࣛࢵࣉࡋ࡚࠾ࡾࠊ⅖㛵㐃 P51$ ࡢ ㄪ⠇ᶵᵓࡢ୍➃ࢆ᫂ࡽࡍࡿࡇࡀฟ᮶ࡓࠋࡲࡓࠊ 1%3 ࢆጞࡵ 5HJQDVH ࣇ࣑࣮ࣜศᏊḞᦆ࣐࢘ࢫ ࢆ⏝࠸ࡓ 7UDQVFULSWRPH ゎᯒࢆ⾜ࡗ࡚࠾ࡾࠊࡇࡢศ Ꮚ⩌ࡢචㄪ⠇ᶵᵓゎ᫂ᐤࡍࡿࡇࡀᮇᚅࡉࢀ ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 51$ ⤖ྜ⺮ⓑ㉁ࡢᶵ⬟ࡸࡑࡢᶆⓗ P51$ ࢆ᫂ࡽࡍ ࡿࡣࠊ⥙⨶ⓗ࡞ゎᯒࡀᚲせྍḞ࡛࠶ࡿࠋ࠼ࡤࠊ ᅇᨭࡋ࡚࠸ࡓࡔ࠸ࡓ 5HJQDVH Ḟᦆ 7 ⣽⬊࠾ ࡅࡿ 7UDQVFULSWRPH ゎᯒ࡛ࡣࠊḞᦆ⣽⬊࡛≉␗ⓗ Ⓨ⌧ୖ᪼ࡍࡿ P51$ ࡋ࡚ࠊF5HO ࡞ࡇࢀࡲ࡛⪃࠼ ࡚ࡇ࡞ࡗࡓ᪂つ㑇ఏᏊࢆྠᐃࡍࡿࡇࡀฟ᮶ࡓࠋ ࡇࢀࡣࠊḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮ࢆ⏝࠸ࡓ⥙⨶ⓗゎᯒ ࢆ⾜ࢃࡎࡋ࡚ࡣᚓࡿࡇࡢฟ᮶࡞ࡗࡓ▱ぢ࡛࠶ࡿࠋ ࡇࡢ▱ぢࡼࡾࠊ5HJQDVH ࡼࡾ㌿ᅉᏊࡢ୍ࡘ F5HO ࡀไᚚࡉࢀࠊࡑࡢ⤖ᯝࡋ࡚ 7 ⣽⬊άᛶࢆไ ᚚࡍࡿ࠸࠺᪂ࡋ࠸ࣔࢹࣝࢆᚓࡿࡇࡀฟ᮶ࡓࠋ ࡲࡓࠊ ᖺ ᖺᗘࡢᨭࡼࡾ 5HJQDVH 5RTXLQ ࡢඹ㏻ᶆⓗ P51$ ࡢᏳᐃᛶไᚚࡢ࣓࢝ࢽࢬ࣒ ࢆ᫂ࡽࡋ㸪ࡇࢀࡲ࡛ࡃゎ᫂ࡉࢀ࡚࠸࡞࠸ྠ୍ 51$ ㄆ㆑㓄ิࢆ⏝࠸ࡓ」ᩘࡢ㌿ᚋㄪ⠇ᅉᏊࡼࡿ ཝᐦ࡞ 51$ Ᏻᐃᛶไᚚࡢゎ᫂ᒎ㛤ࡉࡏࡿࡇࡀฟ ᮶ࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 8HKDWD7HWDO&HOO 0LQR7HWDO&HOO - 413 - බເᨭㄢ㢟㸸2013 ᖺᗘ ࡞ࡐ⥺ࡀᕧᮌࢆᯤࡽࡍࡢ̿㑇ఏᏛⓗᡭἲࢆ⏝࠸ࡓ࣐ࢶᮦ⥺ཎ࣓࢝ࢽࢬ࣒ࡢゎ᫂ ᣮᡓⓗⴌⱆ◊✲ ➉ෆ ♸Ꮚ࣭ி㒔ᏛᏛ㝔㎰Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ཎ⥺࣐ࢶࣀࢨࢭࣥࢳࣗ࢘ࡀఏ᪻ࡼࡗ ࡚࣐ࢶᒓᶞᮌᑟධࡉࢀࡿࡇ࡛㉳ࡇࡿ࣐ࢶᮦ⥺ ࡣࠊⓑ◁㟷ᯇࢆ᧦ࡍࡿ᪥ᮏྂ᮶ࡢᬒほࢆቯ⁛≧ែ ㏣࠸㎸ࡳࠊ࡞࠾ᣑࢆ⥆ࡅ࡚࠸ࡿࠋᮏࡢཎ ᶵᵓࡀఱ㉳ᅉࡍࡿࡢࡣ࠸ࡲࡔゎ᫂ࡉࢀ࡚࠾ࡽࡎࠊ ᑐ⒪ἲ౫Ꮡࡏࡊࡿࢆᚓ࡞࠸⌧⾜ࡢ㜵㝖⟇ࡣ᭷ຠ ᶵ⬟ࡋ࡚࠸࡞࠸ࠋᮏ◊✲࡛ࡣࠊ࠸ࡲࡔㅦࡢከ࠸ᮏ ཎ࣓࢝ࢽࢬ࣒ࡢᐜゎ᫂ࢆ┠ⓗࡋ࡚ࠊ⊂⮬ సฟࡋࡓᮏ⥺ࡢ⤌࠼㏆⣔ࢆᮦᩱࡋࡓࢤࣀ࣒ ࣡ࢻ࡞ሷᇶከᆺゎᯒ⾲⌧⣔ゎᯒࢆ⤌ࡳྜࢃࡏ ࡿࡇࡼࡾࠊ࣐ࢶࣀࢨࢭࣥࢳࣗ࢘ࡢ᰾ࢤࣀ࣒ୖ ࠶ࡿணࡉࢀࡿཎᛶỴᐃᅉᏊ㸦ᐩᶔࡽ 2009㸧 ࡢ≉ᐃࢆ┠ᣦࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࣐ࢶࣀࢨࢭࣥࢳࣗ࢘⤌࠼㏆⣔ 17 ⣔⤫ཬࡧ ࡑࢀࡽࡢぶ⣔⤫࡛࠶ࡿ㏆⣔ 2 ⣔⤫ࡢィ 19 ⣔⤫ࡘ ࠸࡚ࠊRAD-tag ࡢࣜࣇࣞࣥࢫࢤࣀ࣒ࡢ࣐ࢵࣆࣥ ࢢࡑࡢゎᯒ㝿ࡋ࡚ᨭࢆཷࡅࡓࠋࣜࣇࣞࣥࢫ ࢤࣀ࣒ࡣ GeneDB ࢙࢘ࣈࢧࢺୖࡢ࣐ࢶࣀࢨࢭ ࣥࢳࣗ࢘ Ka4C1 line ࢻࣛࣇࢺࢤࣀ࣒ࢆ⏝ࡋࡓ 㸦http://www.genedb.org/Homepage/Bxylophilus㸧ࠋ ࢤࣀ࣒࣡ࢻ㛵㐃ゎᯒࡼࡾ㔜せᙧ㉁┦㛵ࡀᚓࡽ ࢀࡓ㑇ఏᏊ㡿ᇦࡘ࠸࡚ࡢࣀࢸ࣮ࢩࣙࣥཬࡧ GO ゎᯒࢆሗゎᯒᨭࡢࡶ࡛⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᅜෆእ࡛῝้࡞⿕ᐖࢆཬࡰࡋ⥆ࡅࡿ᳃ᯘὶ⾜࡛ ࠶ࡿ࣐ࢶᮦ⥺ࡢཎ⥺࣐ࢶࣀࢨࢭࣥࢳࣗ࢘ ࠾࠸࡚ࠊᐟࡢⓎཬࡧ⿕ᐖᣑᐦ᥋㛵ࡍ ࡿ㔜せᙧ㉁ࡋ࡚ࠊཎຊࠊቑṪຊࠊఏ᪻ࡢ ౽ຊࡢ 3 ࡘࡀᣲࡆࡽࢀࡿࠋᮏ✀ࡣ 2011 ᖺࢻࣛࣇ ࢺࢤࣀ࣒ሗࡀබ㛤ࡉࢀࡓࡀ㸦Kikuchi et al., PLOS Pathogens, 2011㸧࠸ࡲࡔศᏊᇶ┙ࡢᩚഛ㏵ୖ࠶ࡾࠊ ࡇࢀࡽࡢ㔜せᙧ㉁ࢆไᚚࡍࡿ㑇ఏᏊ㸦⩌㸧ࡘ࠸࡚ ࡶࡇࢀࡲ࡛ᮍゎ࡛᫂࠶ࡗࡓࠋᮏ◊✲࡛ࡣྂ㑇ఏᏛ ⓗ࡞ࣉ࣮ࣟࢳࡋ࡚ࠊ⤌࠼㏆⣔ࢆ᪂ࡓసฟ ࡋ࡚⥺ᮦᩱ౪ࡋࠊRAD-seq ἲࡼࡿࢤࣀ࣒ゎᯒ ࢆ⾜࠺ࡇ࡛ཎᛶ㛵㐃㑇ఏᏊ⩌ࡢ᥎ᐃࢆ⾜ࡗࡓࠋ ࡇࢀࡼࡾࠊୖグࡢྛᙧ㉁ᑐࡋ࡚㧗࠸┦㛵ࢆᣢࡘ 3 㡿ᇦࢆࢻࣛࣇࢺࢤࣀ࣒ୖぢฟࡋࡓࠋ ⥆࠸࡚ࠊྛᙧ㉁㧗࠸┦㛵ࢆᣢࡘ 3 㡿ᇦୖᏑᅾ ࡍࡿ㑇ఏᏊ⩌ࢆゎᯒࡍࡿࡇ࡛ࠊࡢࡼ࠺࡞㑇ఏᏊ ࡀࡑࢀࡒࢀࡢᙧ㉁㛵ࡋ࡚࠸ࡿࢆ᥎ᐃࡋࡓࠋ ཎຊ≉␗ⓗ࡞┦㛵ࢆࡶࡘ㡿ᇦ࠾࠸࡚ࡣࠊ࣒࣊࣌ࣝ ࢜࢟ࢩࢲ࣮ࢮࡸ CYP ➼ࡢ௦ㅰ㛵ࡍࡿ㑇ఏᏊࡀ ᳨ฟࡉࢀࡓࠋཎຊ㧗࠸┦㛵ࢆ♧ࡋࡓ㡿ᇦࡣ࠸ࡎ ࢀࡶቑṪຊ㧗࠸┦㛵ࡀㄆࡵࡽࢀࠊቑṪຊࡣཎຊ ࡗ࡚ษࡾ㞳ࡏ࡞࠸ᙧ㉁࡛࠶ࡿࡇࡀ♧၀ࡉࢀࡓࠋ ୧ᙧ㉁㧗࠸┦㛵ࢆ♧ࡋࡓ㡿ᇦࡣࠊⓎ⏕ࡸᡂ㛗 㛵ࡍࡿࢩࢢࢼࣝ࣌ࣉࢳࢲ࣮ࢮ㑇ఏᏊࡀ᳨ฟࡉࢀࡓࠋ ࡑࡢࠊቑṪຊ౽ຊࡢ୧᪉┦㛵ࢆࡶࡘ㡿ᇦ࡛ ࡣෆ⬇ⴥᙧᡂࡸᙧែᙧᡂ㛵ࡍࡿ daf-1 㑇ఏᏊࡀ ᳨ฟࡉࢀࠊࡑࢀࡒࢀࡢᙧ㉁ࡁࡃ㛵ࡋ࡚࠸ࡿࡇ ࡀ᥎ᐹࡉࢀࡓࠋ ᚋࡣࠊᮏ◊✲࡛ぢฟࡉࢀࡓྛ㔜せᙧ㉁㛵㑇ఏ Ꮚࡢೃ⿵ࡘ࠸࡚ࠊᘬࡁ⥆ࡁ㑇ఏᏊⓎ⌧ゎᯒࡸ㑇ఏ Ꮚࣀࢵࢡࢲ࢘ࣥཬࡧࣀࢵࢡ࢘ࢺ➼ࠊヲ⣽࡞ᶵ⬟ゎ ᯒࢆ⾜࠸ࠊཎᛶࡢ㛵ࡢ᭷↓ࢆ᳨ドࡋࡓ࠸ࠋ ୖグࡢࡼ࠺࡞ᡂᯝࡀᚓࡽࢀࡓ୍᪉࡛ࠊᮏ◊✲ࡢ⢭ ᗘ࠾࠸࡚⏕≀ᮦᩱ࡛࠶ࡿ⤌࠼㏆⣔ࡢ⣔⤫ᩘࠊ ࡞ࡽࡧసฟ࠾ࡅࡿ⤌࠼㢖ᗘࡢపࡉࡢᨵၿࡀ ㄢ㢟࡞ࡗࡓࠋᮏ✀ࡣ㞤㞝␗ಶయ࡛᭷ᛶ⏕Ṫࢆ⾜࠺ Ⅼ࡛ࠊྠࡌࡃ⥺ᙧື≀㛛ᒓࡍࡿࣔࢹࣝ⏕≀ࡢ Caenorhabditis elegans ẚ㍑ࡋ࡚⤌࠼㏆⣔స ฟࡢ㞴᫆ᗘࡣ㧗࠸ゝ࠼ࡿࡀࠊᮏ◊✲ࡢࡼ࠺࡞㑇ఏ Ꮫⓗࣉ࣮ࣟࢳࢆヨࡳࡿ㝿ࡣ᪂ࡓ࡞⤌࠼㏆⣔ ࡢసฟࡔࡅ࡛࡞ࡃࠊࡑࡢసฟἲࡘ࠸࡚ࡶᨵⰋࡢవ ᆅࡀ࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ࡢᵓẁ㝵࠾࠸࡚ࡣࠊ࣐ࢶࣀࢨࢭࣥࢳ ࣗ࢘⤌࠼㏆⣔ࡢከᆺゎᯒ⏝ࡍࡿ DNA ࣐࣮ ࢝ ࣮ ࡋ ࡚ AFLP 㸦 amplified fragment length polymorphism㸧࣐࣮࣮࢝ཬࡧ࣐ࢡࣟࢧࢸࣛࢺ࣐ ࣮࣮࢝㸦SSR; simple sequence repeat㸧ࢆᐃࡋ࡚ ணഛᐇ㦂ࢆ⾜ࡗ࡚࠸ࡓࡀࠊ࠸ࡎࢀࡶࢤࣀ࣒ࡢ㛵㐃 ࡅࡸከᆺ᳨ฟ⬟ࡢⅬ࡛᭱㐺ࡣゝ࠼࡞ࡗࡓࠋ ᅇࠊࢤࣀ࣒ᨭ㸦ሗゎᯒᨭ㸧ࢆཷࡅࡽࢀࡿࡇ ࡞ࡗࡓࡇ࡛ࠊࡼࡾ⢭⦓࡞ゎᯒࡀྍ⬟࡞ࡿ SNP 㸦single nucleotide polymorphism㸧࣐࣮࣮࢝ࡢ㛤Ⓨ ╔ᡭࡍࡿࡇࡀ࡛ࡁࠊゎᯒせࡍࡿ㛫ࡶᖜ ▷⦰࡛ࡁࡓࠋࡶࡶ᧯సᐇ㦂ࢆయࡍࡿ࢙࢘ࢵ ࢺ࡞◊✲ࢆ⾜ࡗ࡚ࡁࡓᙜ◊✲ᐊ࡛ࠊࢤࣀ࣒ሗゎᯒ యࡢࢻࣛ࡞◊✲ࢆ㐙⾜ࡍࡿࡇࡣࠊᨭ࡞ࡋ ࡣྍ⬟࡛࠶ࡗࡓ⪃࠼ࡽࢀࡿࠋ ࡲࡓࠊᅇࡢᨭࢆ㏻ࡋ࡚ࠊᙜ◊✲ᐊ࠾ࡅࡿࢤ ࣀ࣒ሗゎᯒ⎔ቃ㸦ࣁ࣮ࢻ㠃㸧ࡢᩚഛࠊ࡞ࡽࡧ Ꮫ⏕ࡢゎᯒᢏ⾡ࡢ⩦ᚓࡀࢫ࣒࣮ࢬ࡞ࡗࡓࠋࡉࡽ ࠊᮏ◊✲௨እࡢࣉࣟࢪ࢙ࢡࢺ࠾࠸࡚ࡶࠊ◊✲ィ ⏬ࢆ❧࡚ࡿୖ࡛ࢤࣀ࣒ሗゎᯒࢆ⤌ࡳ㎸ࡴ࡞ࣉ ࣮ࣟࢳࡢ㑅ᢥᖜࡀ᱁ẁᗈࡃ࡞ࡗࡓࠋࡇࡢሙࢆࡾ ࡚㛵ಀ⪅ࡢࡳ࡞ࡉࡲឤㅰࡋࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ヱᙜ࡞ࡋ - 414 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࣈ࣮࣒ࣝ㑇ఏᏊḞᦆ '1$ 㔜㙐ษ᩿ࡢ⤌ࡳྜࢃࡏࡼࡿࢤࣀ࣒Ᏻᐃᛶࡢゎᯒ ᪂Ꮫ⾡㡿ᇦ◊✲ ➉⏣ ₶࣭㜰ᏛᏛ㝔་Ꮫ⣔◊✲⛉ sample based detection of structural variation) 㸺⫼ᬒ࣭┠ⓗ㸼 ⭘⒆ࡀ㧗㢖ᗘฟ⌧ࡍࡿࣄࢺࣈ࣮࣒ࣝೃ⩌ࡢཎ ྡࡅࡓ᪂つࢤࣀ࣒ᵓ㐀ኚ␗ゎᯒࢶ࣮ࣝࢆඹྠ㛤Ⓨ ᅉ㑇ఏᏊ࡛࠶ࡿࣄࢺࣈ࣮࣒ࣝ㑇ఏᏊ㸦BLM 㑇ఏᏊ㸧 ࡋࡓ (Yamagata K et al. in press)ࠋࡑࡢࢶ࣮ࣝࢆ ࡢⓎ⌧ࢆᢚไࡍࡿࠊయ⣽⬊࡛࠶ࡗ࡚ࡶᰁⰍయ㛫ࡢ ࡗࡓゎᯒ⤖ᯝࠊᨺᑕ⥺↷ᑕ⩌࡛ࣈ࣮࣒ࣝ㑇ఏᏊ࢜ࣇ ᕪ㢖ᗘࡀⴭࡋࡃୖ᪼ࡋࠊ∦ࣞࣝࡔࡅኚ␗ࡀ࠶ ࡢ㸯ࢡ࣮ࣟࣥ࠾࠸࡚ࠊᰁⰍయࡢࢸ࣓ࣟ㏆ഐ㡿ᇦ ࡿ⣽⬊ࡀ୧ࣞࣝኚ␗ࢆ᭷ࡍࡿ⣽⬊㧗㢖ᗘ㌿ ࠾࠸࡚ᵓᡂࡀ㞟୰ⓗ㉳ࡁ࡚࠸ࡿࡀほᐹࡉࢀ ࡍࡿࠋࣈ࣮࣒ࣝ㑇ఏᏊḞᦆ≧ែࡢࢤࣀ࣒ኚࢆゎ ࡓࠋࡇࢀࡣࠊࣄࢺࡢ࢞ࣥࢤࣀ࣒࡛ሗ࿌ࡉࢀ࡚࠸ࡿ ᯒࡍࡿࡓࡵࠊࣈ࣮࣒ࣝ㑇ఏᏊࢆ୍ⓗ࢜ࣇ࡛ Chromothripsis(Stephens PJ et al., Cell.2011)㢮 ࡁࡿ F㸯(B6×129) ࢱࣉࡢ࣐࢘ࢫ ES ⣽⬊ࢆᶞ❧ࡋࠊ ఝࡋ࡚࠸ࡓࠋࡉࡽࠊࡑࡢࢡ࣮ࣟࣥࡽࠊ␗࡞ࡗࡓ ࣊ࢸࣟ᥋ྜᛶࡢᾘኻ㸦Loss of Heterozygosity: LOH㸧 ᵓᡂࢆ᭷ࡍࡿࢧࣈࢡ࣮ࣟࣥࡀὴ⏕ࡋ࡚࠸ࡿࡀ ࡀ⏕ࡌࡓ⣽⬊ࢆ᳨ฟ࡛ࡁࡿࢩࢫࢸ࣒ࢆᵓ⠏ࡋࡓࠋ arrayCGH (Comparative genomic hybridization)࡞ ࣈ࣮࣒ࣝ㑇ఏᏊḞᦆ≧ែ DNA 㔜㙐ษ᩿ࢆ⤌ࡳ ࡢᡭἲࡼࡾ☜ㄆ࡛ࡁࡓࠋ ྜࢃࡏ࡚ࠊAprt 㑇ఏᏊᗙ㛵ࡋ LOH ࡀ㉳ࡁࡓࢡࣟ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣮ࣥࢆ㑅ᢥࡋࠊゎᯒࡋࡓࠋ࡞࠾ࠊDNA 㔜㙐ษ᩿ࡣ ࣄ ࢺ ࡢ ࢞ ࣥ ࢤ ࣀ ࣒ ࡛ ሗ ࿌ ࡉ ࢀ ࡚ ࠸ ࡿ Aprt 㑇ఏᏊᗙ㏆ഐ≉␗ⓗᑟධࡋࡓࠋࡑࡢ⤖ᯝࠊ Chromothripsis ࡣࠊྠ⏕ࡌࡓ」ᩘࡢࢤࣀ࣒᩿∦ Aprt 㑇ఏᏊᗙ࿘㎶ࠊ」㞧࡞ᵓᡂࡀ㉳ࡁ࡚࠸ࡿࡇ ࡀ⤖ྜࡋࠊᵓᡂ㡿ᇦࢆゎᯒࡍࡿࢤࣀ࣒ࢥࣆ࣮ ࡀุ᫂ࡋࡓ㸦Yamanishi A et al. Genome Res 23: ᩘࡣࠊኚῶࡌ࡚࠸ࡿࡀ▱ࡽࢀ࡚࠸ࡿࠋࡘࡲࡾࠊ 1462-1473, 2013㸧ࠋ 」〇ᶵᵓࡣ㛵ࡋ࡚࠸࡞࠸ࡇࡀࠊ♧၀ࡉࢀ࡚࠸ࡿࠋ ࡑࡇ࡛ᮏᨭ࠾࠸࡚ࡣࠊᨺᑕ⥺↷ᑕࡼࡾࢤࣀ࣒ ࡇࢁࡀࠊᨺᑕ⥺↷ᑕ⩌ࠊࣈ࣮࣒ࣝ㑇ఏᏊ࢜ࣇࡢࢤ యࢃࡓࡗ࡚ DNA 㔜㙐ษ᩿ࢆᑟධࡋࠊྠᵝࡢ ࣀ࣒ࢆ arrayCGH ࡛ゎᯒࡋ࡚ࡳࡿࠊᵓᡂ㞟✚㡿 ᵓᡂࡀ㉳ࡁ࡚࠸ࡿ࠺ࢆ᳨ウࡍࡿࡇࡋࡓࠋ ᇦ࠾࠸࡚ࠊࢥࣆ࣮ᩘࡀቑຍࡋ࡚࠸ࡿࡀุ᫂ࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࡇࡢࡇࡣࠊࣈ࣮࣒ࣝ㑇ఏᏊࡀ࢜ࣇࡢぢࡽࢀࡿ ᨺᑕ⥺↷ᑕ⩌ࡽࠊࣈ࣮࣒ࣝ㑇ఏᏊ࢜ࣥ㸯ࢡ࣮ࣟࣥࠊ ࢤࣀ࣒Ᏻᐃ㉳ࡁࡿࢤࣀ࣒ᵓᡂࡣࠊ」〇ᶵ ࣈ࣮࣒ࣝ㑇ఏᏊ࢜ࣇ㸯ࢡ࣮ࣟࣥࠊᨺᑕ⥺㠀↷ᑕ⩌ ᵓࡀ㛵ࡋ࡚࠸ࡿࡀ♧၀ࡉࢀࠊ⭘⒆Ⓨᶵᵓࡢ ࡽࠊࣈ࣮࣒ࣝ㑇ఏᏊ࢜ࣥ㸯ࢡ࣮ࣟࣥࠊࣈ࣮࣒ࣝ㑇ఏ ᪂ࡓ࡞࣓࢝ࢽࢬ࣒ࢆ♧ࡋ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿᛮࢃ Ꮚ ࢜ ࣇ 㸰 ࢡ ࣟ ࣮ ࣥ ࡼ ࡾ ࢤ ࣀ ࣒ DNA ࢆ ᢳ ฟ ࡋ ࠊ ࢀࡿࠋ illumina HiSeq 2500 ࡼࡿ paired-end sequencing (500-bp size range, 100-bp read length, 30 depth) 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡼࡾࢤࣀ࣒ࢩ࣮ࢡ࢚ࣥࢫࢆ౫㢗ࡋࡓࠋ Yamagata K. et al., Nucleic Acids Res., in press 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᚓࡽࢀࡓࢹ࣮ࢱࡽࢤࣀ࣒ᵓᡂࡀ㉳ࡁࡓᛮࢃࢀ ࡿ㡿ᇦࢆ≉ᐃࡍࡿࡓࡵࠊ℩ࠎ₶௵◊✲ဨ㸦⏘ᴗ ᢏ⾡⥲ྜ◊✲ᡤ㸧ࡽࡶࠊCOSMOS (Control - 415 - බເᨭㄢ㢟㸸2013 ᖺᗘ ඹ⏕యឤᰁࡼࡾኚືࡍࡿඹ⏕㑇ఏᏊไᚚࢿࢵࢺ࣮࣡ࢡࡢゎ᫂ ⱝᡭ◊✲㸦㹀㸧 Ṋ⏣ ┤ஓ࣭ᇶ♏⏕≀Ꮫ◊✲ᡤඹ⏕ࢩࢫࢸ࣒◊✲㒊㛛 㸺⫼ᬒ࣭┠ⓗ㸼 ᳜≀ࡣ⳦᰿⳦ࡸ᰿⢏⳦࡞ࡢᚤ⏕≀ࡢඹ⏕ࡼࡾ ࣭ࣜࣥ❅⣲※ࢆᚓࡿࡇࡀ࡛ࡁࠊ᳜≀ࡢ⎔ቃ㐺ᛂࡸ ⏕⫱ಁ㐍ࡁࡃ㈉⊩ࡋ࡚࠸ࡿࠋᮏ◊✲࡛ࡣࠊࡇࡢ ඹ⏕ᰤ㣴౪⤥ຠᯝࢆࡶࡓࡽࡍඹ⏕ᚤ⏕≀ࡢᐟ᳜≀ ࡢឤᰁᶵᵓࡘ࠸࡚ࡢゎᯒࢆ⾜࠸ࠊ⏕≀㛫┦స ⏝ࡼࡾ␗✀⏕≀ࢆయෆཷᐜࡍࡿศᏊᶵᵓࡢゎ᫂ ࢆ┠ⓗࡋ࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࣐࣓⛉᳜≀࣑ࣖࢥࢢࢧ(Lotus japonicus)࡛ྠᐃࡋࡓ ࣮ࣂࢫ࣮࢟ࣗࣛ⳦᰿㸦AM㸧ඹ⏕࣭᰿⢏ඹ⏕ኚ␗య ࢆ⏝࠸࡚ࠊࡇࢀࡽࡢኚ␗యࡀ♧ࡍ AM ⳦࣭᰿⢏⳦ࡢ ឤᰁ␗ᖖࢆᫎࡋࡓ㑇ఏᏊⓎ⌧ࣉࣟࣇࣝࠊ㠀ឤ ᰁࡢ᳜≀యࡢ㑇ఏᏊⓎ⌧ࣉࣟࣇࣝࡢゎᯒࢆ RNA-Seq ࡼࡾ⾜ࡗࡓࠋࡲࡓࠊඹ⏕ࢩࢢࢼࣝࢆᜏᖖ ⓗάᛶࡉࡏࡿࡇࡀ࡛ࡁࡿᶵ⬟⋓ᚓᆺඹ⏕ࢩࢢ ࢼࣝᅉᏊ gain of function CCaMK ࢆ㐣Ⓨ⌧ࡉࡏ ࡓ᰿࠾ࡅࡿⓎ⌧ࣉࣟࣇࣝࡶྲྀᚓࡋࠊඹ⏕ኚ␗ య࡛ᚓࡽࢀࡓࣉࣟࣇࣝࡢẚ㍑⏝࠸ࡓࠋ ゎᯒࢆ⾜ࡗࡓኚ␗యࡢ࠺ࡕࠊ㌿ไᚚ㛵ࢃࡿ⪃ ࠼ࡽࢀࡿ NSP1 RAMI1 ࡘ࠸࡚ࠊc-myc tag ࡛ᶆ ㆑ࡋࡓࢱࣥࣃࢡ㉁ࢆචỿ㝆ࠊDNA ㄪᩚࢆ⾜ࡗࡓࢧ ࣥࣉࣝࡘ࠸࡚ࠊḟୡ௦ࢩ࣮ࢣࣥࢫࡼࡿ Chip-Seq ゎᯒࢆ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣑ࣖࢥࢢࢧ࡛ྠᐃࡋࡓ AM ⳦࣭᰿⢏⳦ࡢ୧ឤᰁ㐣⛬ ࡀ୰㏵࡛Ṇࡍࡿඹ⏕ኚ␗య cerberus ࢆ⏝࠸࡚ࠊࡇ ࢀࡽࡢኚ␗యࡀ♧ࡍ⳦᰿࣭᰿⢏ඹ⏕యࡢឤᰁ␗ᖖࢆ ᫎࡋࡓ㑇ఏᏊⓎ⌧ࣉࣟࣇࣝࢆྲྀᚓ࣭ゎᯒࡋࡓࠋ ࡑࡢ⤖ᯝࠊࡇࢀࡽࡢኚ␗⾲⌧ᆺࢆᘬࡁ㉳ࡇࡍ⪃࠼ ࡽࢀࡿ㑇ఏᏊⓎ⌧ࡢኚࡋ࡚ࠊ⣽⬊ቨྜᡂ㓝⣲㑇 ఏᏊ⩌ࡢⓎ⌧ኚືࡀࡳࡽࢀࡓࡇࡽࠊࡇࡢ㑇ఏᏊ ⩌ὀ┠ࡋ࡚◊✲ࢆ⾜࠺ࡇࡋࡓࠋcerberus ኚ␗ య ࡛ ࡣ ឤ ᰁ ᰿ ࠾ ࠸ ࡚ Xyloglucan endotrans -glucosylase / hydrolases (XTHs) ࡀ㔝⏕ᆺẚ ᭷ពపୗࡋ࡚࠸ࡓࠋࡲࡓࠊcerberus ኚ␗య࡛ࡣ㠀 ឤᰁ᰿࠾࠸࡚ࡶ XTH 㑇ఏᏊ⩌ࡢㄏᑟࡀࡳࡽࢀࡓࠋ ࡇࢀࡽࡢ⣽⬊ቨᵓ⠏㓝⣲ XTH ࡣࠊ⣽⬊ቨ୰ࡢࢭࣝ ࣮ࣟࢫ࢟ࢩࣟࢢࣝ࢝ࣥࡢᯫᶫࢆ⧅ࡂኚ࠼ࡿࡇ࡛ ᳜≀ࡢ⣽⬊ቨࡢᵓᡂࢆ⾜࠸ࠊ◳ᗘࢆኚࡉࡏࡿ㓝 ⣲ࡢ㸯ࡘ࡛࠶ࡿࠋඹ⏕⳦࡛࠶ࡿ AM ⳦ࡣ᰿ࡢ⣽⬊㛫 㝽ࢆ⦭࠺ࡼ࠺ఙ㛗ࡋࠊᐟ᰿ෆᗈࡀࡗ࡚࠸ࡃࡇ ࡽࠊ⣽⬊ቨࡢ◳ᗘឤᰁࡢ㛵㐃ࡀ♧၀ࡉࢀࡿࠋ ࡲࡓࠊXyloglucan ≉␗ⓗ࡞ᢠయࡼࡿචᰁⰍࡼ ࡗ࡚ࠊ⳦᰿⳦ࡢ࿘㎶ࡢ⣽⬊ቨ࡛ Xyloglucan ࡢ✚ࡀ ࡳࡽࢀࠊඹ⏕⳦ឤᰁ⣽⬊ቨࡢ㛵㐃ࡀ♧၀ࡉࢀࡓࠋ ࡉࡽࠊcerberus ኚ␗యࡢ㠀ឤᰁ≧ែ࠾ࡅࡿ⣽⬊ ቨᡂศࢆㄪࡿࠊXyloglucan ࡢᵓᡂ༢⢾࡛࠶ࡿ Xylose ࡢྵ㔞ࡀపୗࡋ࡚࠸ࡓࠋࡇࡢࡼ࠺ࠊcerberus ኚ␗య࡛㑇ఏᏊⓎ⌧ࡸ⣽⬊ቨᡂศ␗ᖖࡀࡳࡽࢀࡓ ࡇࡽࠊAM ⳦ࡢ⳦⣒ఙ㛗ᙳ㡪ࢆ࠼ࡿኚ␗⾲ ⌧ᆺࡢ┦㛵ࡀ♧၀ࡉࢀࠊCERBERUS ࡼࡿ⣽⬊ ቨ◳ᗘࡢไᚚࡀ AM ⳦⳦⣒ఙ㛗ࢆไᚚࡋ࡚࠸ࡿྍ⬟ ᛶࡘ࠸࡚ࡢ◊✲ࢆ㐍ࡵ࡚࠸ࡿࠋ ูࡢࣉ࣮ࣟࢳࡋ࡚ࠊAM ඹ⏕ㄏᑟࡉࢀࡿ ࣉࣟࢸ࣮ࢮ㑇ఏᏊ SbtM1 ࡢࣉ࣮ࣟࣔࢱ࣮ࡽ⳦᰿ ඹ⏕≉␗ⓗࢩࢫ㡿ᇦࢆ≉ᐃࡋࠊ㓝ẕ㸯ࣁࣈࣜࢵࢻ ⣔ࡼࡿゎᯒࡽࠊࡇࡢࢩࢫ㓄ิ⤖ྜࡍࡿࡇ࡛ ⳦᰿ඹ⏕㑇ఏᏊࡢ㌿ไᚚࢆ⾜࠺⪃࠼ࡽࢀࡿࢺࣛ ࣥࢫᅉᏊೃ⿵ࢆྠᐃࡋࡓࠋࡇࡢ rami1 ◚ቯᰴࡢࢺࣛ ࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࡽࠊRAMI1 Ⓨ⌧ࡼࡗ࡚ Ⓨ⌧ไᚚᖸ΅ࢆ࠺ࡅࡿඹ⏕㑇ఏᏊ⩌ࡢ᥈⣴ࢆ⾜ࡗ ࡓࠋࡑࡢ⤖ᯝࠊrami1 ◚ቯᰴ࡛ࡣࣇࣛ࣎ࣀࢻྜᡂࠊ ගྜᡂ㛵ࢃࡿ㑇ఏᏊ⩌ࡢⓎ⌧ࡀ㔝⏕ᆺẚ࡚᭷ ពపୗࡋ࡚࠸ࡓࠋࣇࣛ࣎ࣀࢻྜᡂ㑇ఏᏊࡣ AM ඹ⏕୰Ⓨ⌧ࡀኚືࡍࡿ㑇ఏᏊ࡛࠶ࡾࠊඹ⏕࠾ࡅ ࡿᶵ⬟ࡶ▱ࡽࢀ࡚࠸ࡿࠋࡑࡢࡓࡵࠊrami1 ◚ቯᰴ ࠾ࡅࡿ⳦᰿⳦ឤᰁ⋡ࡢୖ᪼ࡢ㛵㐃ࡀ♧၀ࡉࢀࡓࠋ ࡇࢀࡽࡢ㑇ఏᏊࡢࣉ࣮ࣟࣔࢱ࣮㡿ᇦ RAMI1 ࡀ┤ ᥋⤖ྜࡍࡿࡇ࡛Ⓨ⌧ไᚚࢆ⾜ࡗ࡚࠸ࡿྍ⬟ᛶ ࡘ࠸࡚ࠊChip-Seq ࡼࡿゎᯒࢆヨࡳࡓࡀࠊࡇࢀࡽࡢ 㑇ఏᏊᑐࡋ࡚᫂☜࡞⤖ྜࢆ♧ࡍ⤖ᯝࡣᚓࡽࢀ࡞ ࡗࡓࠋࡇࡢᐇ㦂㛵ࡋ࡚ࡣ Chip 㛵ࡍࡿ᮲௳᳨ウࡀ ᚲせ࡛࠶ࡾࠊᚋࡢ◊✲ࡢㄢ㢟࡞ࡿࠋ ᮏ◊✲࡛ࡣࠊඹ⏕ኚ␗యࡢឤᰁ࣭㠀ឤᰁࡢࢺ ࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࡽࠊឤᰁ㐍⾜ࡢྛࢫࢸ࣮ ࢪ࡛㘽࡞ࡿඹ⏕㑇ఏᏊࡶࠊኚ␗㑇ఏᏊࡽ ኚ␗⾲⌧ᆺ⮳ࡿ㛫ࠊ࠶ࡿ࠸ࡣࡑࢀࡽᐦ᥋㛵ಀ ࡋࡓ㑇ఏᏊⓎ⌧⌧㇟ࢆぢฟࡍࡇࡀ࡛ࡁࡓࠋ≉ 㠀ឤᰁࡢ᳜≀యࡢ≧ែࡀඹ⏕⳦ࡢឤᰁᙳ㡪ࢆ ࠼ࡿࡇࡣࠊᚋࡢඹ⏕ᶵᵓࡢゎᯒࡑࡢ⏝ࢆ⪃ ࠼ࡿୖ࡛㔜せ࡞▱ぢ࡞ࡿ⪃࠼ࡽࢀࡿࠋᚋࡣⓎ ぢࡋࡓඹ⏕㑇ఏᏊ⌧㇟ࡢඹ⏕⳦ឤᰁ࠾ࡅࡿᶵ⬟ ࢆヲ⣽ゎᯒࡋࠊ◊✲ᡂᯝࡋ࡚ࡲࡵ࡚࠸ࡃࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡇࢀࡲ࡛ὀ┠ࡉࢀ࡚࠸࡞ࡗࡓ㑇ఏᏊไᚚᶵᵓࡸ⏕ ⌮⌧㇟ࡀࠊඹ⏕࠾࠸࡚㔜せ࡞ᶵ⬟ࢆᣢࡘࡇࡀ᫂ ࡽ࡞ࡗࡓࠋࡇࡢᡂᯝࡣࠊ⛉◊㈝࠾࠸࡚ࡣࡌࡵ ⾜ࡗࡓḟୡ௦ࢩ࣮ࢣࣥࢫゎᯒࡽ᫂ࡽ࡞ࡗࡓ ᡂᯝᑐࡋ࡚ࠊࢤࣀ࣒ᨭࡼࡾⓎᒎⓗ࠶ࡿ࠸ࡣᣑ ᙇࡋ࡚◊✲ࢆ⾜ࡗࡓ⤖ᯝ࡛࠶ࡿࠋࡑࡢࡓࡵࠊࡇࡢࢤ ࣀ࣒ᨭࢆཷࡅࡿࡇ࡛ࠊᮏ᮶ࡣḟࡢ⛉◊㈝◊✲ࡲ ࡛ᚅࡓ࡞ࡃ࡚ࡣ࠸ࡅ࡞࠸ᡂᯝࢆ࠶ࡆࡿࡇࡀ࡛ࡁࠊ ◊✲ࡢⓎᒎࡁࡃ㈉⊩ࡋࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾࠊᢞ✏‽ഛ୰ - 416 - බເᨭㄢ㢟㸸2010ࠥ2011 ᖺᗘࠊ2013 ᖺᗘ ⤌⧊ࡀࡿ࣐ࢡ࡛ࣟࣟࣂࢫࢺ࡞ࢥࣥࣃ࣮ࢺ࣓ࣥࢺࡢᡂ❧࣭⥔ᣢࡢࣟࢪࢵࢡ ᪂Ꮫ⾡㡿ᇦ◊✲ Ṋ⏣ ὒᖾ࣭ᮾிᏛᏛ㝔⌮Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ㌿ᅉᏊ Zic1(࠾ࡼࡧ Zic4)ࡣయ⠇࠾ࡼࡧయ⠇⏤᮶⤌ ⧊ࡢ⫼ഃ࠾࠸࡚⬇ᮇࡽᡂ㨶ࡲ࡛㛗ᮇ㛫ࢃࡓࡾ Ⓨ⌧ࡋ⥆ࡅࠊయᖿ㒊ࡢ⫼ഃᙧែࢆไᚚࡍࡿࠋࡇࢀࡲ ࡛ࡢᙜ◊✲ᐊ࠾ࡅࡿゎᯒࡽࠊZic1 ࡢⓎ⌧ࡀ⬇ᮇ ࠾࠸࡚ࡣእ㒊ࡽࡢࢩࢢࢼࣝࡼࡾㄪ⠇ࡉࢀࡿࡇ ࡸࠊᡂ㨶࠾࠸࡚ࡣ⣽⬊⮬ᚊⓗ⥔ᣢࡉࢀࡿࡇ ࢆ♧၀ࡍࡿ⤖ᯝࡀᚓࡽࢀ࡚࠸ࡓࠋᡃࠎࡣࠊ࢚ࣆࢪ࢙ ࢿࢸࢵࢡ࡞ࢡ࣐ࣟࢳࣥಟ㣭╔┠ࡍࡿࡇ࡛ Zic1 ࡢ㛗ᮇ㛫ࢃࡓࡿⓎ⌧⥔ᣢࡢศᏊ࣓࢝ࢽࢬ࣒ࢆゎ᫂ ࡍࡿࡇࢆ┠ⓗゎᯒࢆ⾜ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 ձBisulfite seq ࣓ࢲ࢝ᡂ㨶㸦㔝⏕ᆺ࣭యᖿ㒊ࡢ⫼ഃᙧែࡀ⭡ഃ ࡍࡿ Da ኚ␗య㸧ࡢ⫼ഃ/⭡ഃ➽⠇ ࣓ࢲ࢝⛶㨶㸦ཷ⢭ᚋ㸵᪥㸧ࡢ⫼ഃ࠾ࡼࡧ⭡ഃ➽⠇ ղChIP-seq ࣓ࢲ࢝⬊⬇ᮇ࠾ࡼࡧᡂ㨶ࡢ⫼ഃ࣭⭡ഃ➽⠇࠾ࡅ ࡿ ྛ ✀ ࣄ ࢫ ࢺ ࣥ ಟ 㣭 㸦 H3K4me1, H3K4me2, H3K4me3, H3K27me3, H3K27Ac㸧 ࣓ࢲ࢝ᡂ㨶⫢⮚࠾ࡅࡿ H3K4me3ࠊH3K27me3 ࣓ࢲ࢝᱓ᐇ⬇ᮇ࠾ࡅࡿ H3K4me2ࠊH3K27me3 ճRNA seq ゎᯒ ࣓ࢲ࢝ᡂ㨶⫼ഃ࠾ࡼࡧ⭡ഃ➽⫗ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 Zic1 㑇ఏᏊᗙࡣ⬊⬇ᮇ࠾࠸࡚ᕧ࡞ DNA ప࣓ ࢳࣝ㡿ᇦࡀᏑᅾࡋࠊࡑࡢ㡿ᇦᢚไᆺ㸦H3K27me3㸧 ࣄࢫࢺࣥಟ㣭ࡀ㧗ࣞ࣋ࣝ࡞ࡗ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋ୍᪉ᡂ㨶ࡢ➽⠇࠾࠸࡚ࡣࠊZic1 ࡢⓎ⌧ ≧ែ┦㛵ࡋ࡚⫼ഃ࡛ࡣάᛶᆺࣄࢫࢺࣥಟ㣭ࡀ㧗ࡃࠊ ⭡ഃ࡛ࡣᢚไᆺࣄࢫࢺࣥಟ㣭ࡀ㧗࠸࠸࠺⤖ᯝࡀᚓ ࡽࢀࡓࡀࠊDNA ࡢ࣓ࢳࣝ㛵ࡋ࡚ࡣ⫼ഃ➽⠇࠾ ࠸࡚≉␗ⓗ࡞ୖ᪼ࡀ᳨ฟࡉࢀࡓࠋ࣓ࢳࣝࡢୖ᪼ࡋ ࡓ㡿ᇦࡣ⫼ഃ➽⠇࠾࠸࡚ᢚไᆺࣄࢫࢺࣥಟ㣭 㸦H3K27me3㸧ࡀῶᑡࡋࡓ㡿ᇦ୍⮴ࡋ࡚࠾ࡾࠊ⬊ ⬇ᮇ࠾࠸࡚⣙ 23kb ࢃࡓࡗ࡚࠸ࡓ DNA ప࣓ࢳࣝ 㡿ᇦࡀࣉ࣮ࣟࣔࢱ࣮㡿ᇦࡢࡳ⦰ᑠࡋࡓࠋࡇࢀࡽ ࡢ⤖ᯝࡽࠊᡂ㨶࠾࠸࡚ࡣ DNA ࣓ࢳࣝࡀᢚไᆺ ࣄࢫࢺࣥಟ㣭ᣕᢠࡋࠊⓎ⌧ࢆ⥔ᣢࡋ࡚࠸ࡿྍ⬟ᛶ ࡀ♧၀ࡉࢀࡓࠋᡃࠎࡣᡂ㨶࡛ほᐹࡉࢀࡓࢡ࣐ࣟࢳࣥ ≧ែࡀ☜❧ࡍࡿࢱ࣑ࣥࢢࢆ᫂ࡽࡍࡿࡓࡵࠊཷ ⢭ᚋ㸵᪥ࡢ⛶㨶ࢆ⏝࠸࡚ࣄࢫࢺࣥಟ㣭ࢆゎᯒࡋࡓ ࡇࢁࠊࣄࢫࢺࣥಟ㣭ࡣ⛶㨶࠾࠸࡚ࡶᡂ㨶ࡰྠ ᵝࡢࣃࢱ࣮ࣥࢆ♧ࡋࡓࠋࡋࡓࡀࡗ࡚ࣄࢫࢺࣥಟ㣭ࡢ ࣃࢱ࣮ࣥࡣ⬇ᮇ࠾࠸࡚ࡍ࡛☜❧ࡋ࡚࠸ࡿ⪃࠼ ࡽࢀࡿࠋ୍᪉࡛ࠊDNA ࣓ࢳࣝࡢ⫼ഃ≉␗ⓗ࡞ୖ᪼ ࡣほᐹࡉࢀ࡞ࡗࡓࡓࡵࠊDNA ࣓ࢳࣝࡣᏤᚋ ⏕ࡌ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ⬊⬇ᮇ࠾ࡅࡿ Zic1 ࡢࢡ࣐ࣟࢳࣥ≧ែࡀⓎ⏕࡛㔜せ࡞ᙺࢆᯝࡓࡍ ㌿ᅉᏊ≉ᚩⓗ࡞ࡶࡢ࡛࠶ࡗࡓࡓࡵࠊᡃࠎࡣᅇ ♧၀ࡉࢀࡓⓎ⌧ไᚚᶵᵓࡢᬑ㐢ᛶࡘ࠸᳨࡚ウࡋࡓࠋ ࡑࡢ⤖ᯝࠊ⬊⬇ᮇ࠾ࡅࡿᕧ࡞ DNA ప࣓ࢳࣝ㡿 ᇦࡢ࠺ࡕ 16%ࡀᡂ㨶➽⠇࠾࠸࡚ࢧࢬࡀ⦰ᑠࡍࡿ ࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡣࠊ⦰ᑠࢆ ཷࡅ࡞࠸㑇ఏᏊẚ㍑ࡋ࡚ᢚไᆺࣄࢫࢺࣥಟ㣭 㸦H3K27me3㸧ࡢࣞ࣋ࣝࡀ᭷ពῶᑡࡋ࡚࠾ࡾࠊⓎ ⌧ࣞ࣋ࣝࡶ㧗࠸ࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡉࡽࠊᏤ ┤ᚋ࠾ࡅࡿࢤࣀ࣒࣡ࢻࡢ DNA ࣓ࢳࣝゎᯒ ࡢ⤖ᯝࡽࠊᡂ㨶࠾࠸࡚ DNA ࣓ࢳࣝࢆཷࡅࡿ㑇 ఏᏊࡢࢇ࡛Ꮴ┤ᚋࡣ DNA ࣓ࢳࣝࡀ⏕ ࡌ࡚࠸࡞࠸ࡇࡶ᫂ࡽ࡞ࡗࡓࠋࡋࡓࡀࡗ࡚ࠊZic1 ࡢゎᯒࡼࡾ♧၀ࡉࢀࡓࠊᏤᚋ⏕ࡌࡿ DNA ࣓ࢳ ࣝࡼࡿⓎ⌧⥔ᣢࡣⓎ⏕㛵ࢃࡿ㔜せ࡞㌿ᅉᏊ ᬑ㐢ⓗ࡞ไᚚ࡛࠶ࡿྍ⬟ᛶࡀ࠶ࡿࠋᐇ㝿ࡑࡢࡼ ࠺࡞㌿ᅉᏊࡢከࡃࡣ⏕ᾭࢃࡓࡾⓎ⌧ࡍࡿࡇ࡛ ⣽⬊ࡢࢹࣥࢸࢸࡢ⥔ᣢᐤࡍࡿ⪃࠼ࡽ ࢀ࡚࠾ࡾࠊࡑࡢⓎ⌧␗ᖖࡣ⣽⬊ࡢ㐠ࢆኚࡉࡏ࡚ ࡋࡲ࠺ࡇࡽࠊࡑࢀࡽࡢ㌿ᅉᏊࡢⓎ⌧ไᚚᶵᵓ ࡢ⌮ゎࡣᴟࡵ࡚㔜せ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡇࡢ⤖ᯝࡣࠊⓎ⏕㔜せ࡞㌿ᅉᏊࡣከ⬟ᛶ⣽⬊ ࠾࠸࡚ᕧ࡞ HMD ࡼࡗ࡚ᙉᅛᢚไࡉࢀ࡚࠸ࡿ ࡇࢆ♧ࡋ࡚࠸ࡿࠋࡇࢀࡽࡢ㑇ఏᏊ⩌ࡣ⣽⬊ࡢ㐠 Ỵᐃⓗ࡞ᙺࢆᣢࡘࡓࡵࠊᮍศ≧ែࢆ⥔ᣢࡍࡿ ࡓࡵࡣᙉࡃᢚไࡉࢀ࡚࠸ࡿᚲせࡀ࠶ࡿ୍᪉࡛ࠊศ ่⃭ᑐࡋ࡚ࡣ㎿㏿Ⓨ⌧ࡍࡿᚲせࡀ࠶ࡿࠋᮍศ ᛶศ⬟ࡢ୧❧ࡣከ⬟ᛶ⣽⬊࠾ࡅࡿᮏ㉁ⓗ࡞ ≉ᚩ࡛࠶ࡾࠊHMD ࡢࢧࢬࡼࡿᢚไ≧ែࡢไᚚࡣ 㔜せ࡞ᙺࢆᯝࡓࡍ⪃࠼ࡽࢀࡿࠋࡇࢀࡲ࡛ࡢ࢚ࣆ ࢪ࢙ࢿࢸࢵࢡ࡞㑇ఏᏊⓎ⌧ไᚚ㛵ࢃࡿ◊✲ࡢከ ࡃࡣಟ㣭ࡢ✀㢮╔┠ࡋࡓࡶࡢ࡛࠶ࡗࡓࡀࠊᡃࠎࡣ ࢚ࣆࢪ࢙ࢿࢸࢵࢡಟ㣭ࢻ࣓ࣥࡢࢧࢬࡀ㑇ఏᏊ Ⓨ⌧ไᚚࡢ㔜せ࡞せ⣲ࡢ୍ࡘ࡛࠶ࡿࡇࢆ᫂ࡽ ࡍࡿࡇࡀฟ᮶ࡓࠋᚋࠊࡇࡢࡼ࠺࡞ࢡ࣐ࣟࢳࣥಟ 㣭㡿ᇦࡢࢧࢬไᚚࡼࡗ࡚ㄝ࡛᫂ࡁࡿ᪂ࡓ࡞⏕ ⌧㇟ࡢⓎぢࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Nakamura, R. et al., Development 141, 2568-2580 (2014) - 417 - බເᨭㄢ㢟㸸2013 ᖺᗘ ศᏊ㛫ᯫᶫࢆᛂ⏝ࡋࡓ NFATc1 」ྜయゎᯒࡼࡿ㦵࢚ࣆࢪ࢙ࢿࢸࢡࢫࡢᇶ┙ⓗ◊✲ ᇶ┙◊✲ B 㱟㔝୍㑻࣭ᮾ㑥Ꮫబ་⒪ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 (1)◊✲ࡢ⫼ᬒ 㧗㱋♫క࠸㦵⢒㧼࡞ࡢ㦵ᝈᝈ⪅ࡢᛴ ⃭࡞ቑຍࡶࢃࡽࡎࠊࡑࡢ⒪⸆ࡢ㑅ᢥ⫥ࡣỴ ࡋ࡚༑ศࡣゝ࠼࡞࠸ࠋࡑࡢែࡢ୰ᚰ⨨ࡍࡿ 㦵ࣜࣔࢹࣜࣥࢢࡣ㦵ᙧᡂ㦵྾ࡢࣂࣛࣥࢫ࡛ไᚚ ࡉࢀ࡚࠸ࡿࡀࠊࡑࡢᮏ㉁ࡣ㦵ⱆ⣽⬊◚㦵⣽⬊ࡢᶵ ⬟ㄪ⠇࡛࠶ࡿࠋ㦵࢚ࣆࢪ࢙ࢿࢸࢡࢫไᚚࡀḟୡ௦ ᆺ⸆ࢱ࣮ࢤࢵࢺࡋ࡚ᮇᚅࡉࢀࡿ୰࡛ࠊ㦵ⱆ⣽⬊ ◚㦵⣽⬊ࡢࢡ࣐ࣟࢳ࡛ࣥࣞ࣋ࣝࡢไᚚศᏊᶵᵓࡣ ࢇ᫂ࡽࡉࢀ࡚࠸࡞࠸ࠋᡃࠎࡣ RAGE ࣜ࢞ ࣥࢻ AGEs ࢩࢢࢼࣝࡢ㦵௦ㅰᑐࡍࡿᙳ㡪ࢆゎᯒ ࡍࡿ㐣⛬࡛ࠊRAGE-AGEs ࢩࢢࢼࣝࡀࠊ㦵ⱆ⣽⬊ ࡽఱࡽࡢศἪᅉᏊࢆࡋ࡚ࠊ◚㦵⣽⬊ࡢศㄪ⠇ ᶵ⬟ไᚚࡋ࡚࠸ࡿࡇࢆぢฟࡋࡓ (JCB, 2009)ࠋ ᭦ᡃࠎࡢ◊✲ࢳ࣮࣒ࡣࠊ㌿ᅉᏊ」ྜయࡼࡿࢡ ࣐ࣟࢳ࣭࢚ࣥࣆࢪ࢙ࢿࢸࢡࢫࡢᶵ⬟ไᚚࢆࢤࣀ࣒ ࣡ࢻ࡛ゎᯒࡍࡿᡭἲࢆ⊂⮬㛤Ⓨࡋ࡚ࡁࡓ(Cell. 2007)ࠋᮏ◊✲࡛ࡣࠊࡇࢀࡽࡢ▱ぢ࣭᪉ἲㄽࢆ◚㦵⣽ ⬊ศࡢ࣐ࢫࢱ࣮ࣞࢠࣗࣞࢱ࣮࡛࠶ࡿ NFATc1 ᛂ ⏝ࡍࡿࠋNFATc1 (nuclear factor of activated T cells c1) ࡣ ◚ 㦵 ⣽ ⬊ ศ ಁ 㐍 ᅉ Ꮚ RANKL (receptor activator of NF-ț% OLJDQG ࡼࡾ᭱ࡶᙉࡃㄏᑟࡉࢀ ࡿ㌿ᅉᏊ࡛࠶ࡾ (Takayanagi H, et al. 2002 Dev Cell)ࠊࡑࡢࣀࢵࢡ࢘ࢺ࣐࢘ࢫࡣᚰ⮚ᘚ⭷Ḟᦆࢆ యࡍࡿ␗ᖖࡼࡾ⫾ᛶ⮴Ṛ࡛࠶ࡿ(De la pompa JL, et al. 1998 Nature; Ranger AM, et al. 1998 Nature)ࠋ ࡑࡢ㦵≉␗ⓗࣀࢵࢡ࢘ࢺ࣐࢘ࢫ࡛ࡣࠊ◚㦵⣽⬊ศ ࡀᢚไࡉࢀ Osteopetrosis (⌮▼)ࢆ᮶ࡍࡇࡀ ▱ࡽࢀ࡚࠸ࡿ (Asagiri M, et al. 2005 J Exp Med; Winslow MM, et al. 2006 Dev Cell)ࠋࡇࢀࡽࡢᐇࡣࠊ NFATc1 ࡀ◚㦵⣽⬊ศࡢ࣐ࢫࢱ࣮ࣞࢠࣗࣞࢱ࣮ ࡋ࡚㔜せ࡞ศᏊ࡛࠶ࡿࡇࢆ♧ࡋ࡚࠸ࡿࠋ◚㦵⣽⬊ ศࡢࣉࣟࢭࢫࡣࡁࡃศࡅ࡚ࠊ(1)⣽⬊⼥ྜ࣭ከ᰾ ᕧ⣽⬊ࡢᙧᡂ࠸ࡗࡓᙧែኚࡽࠊ(2)㦵⾲㠃ࡢ ྾╔࣭㦵྾❐ࡢᙧᡂ࠸ࡗࡓᶵ⬟Ⓨ⌧ࡢࡓࡵࡢࣉ ࣛࢵࢺࣇ࢛࣮࣒ࡢᙧᡂࠊࡉࡽࠊ(3)HCl ࡸ࢝ࢸࣉࢩ ࣥ K ࡞ࡢ㦵྾ᅉᏊ⩌ࡢྜᡂ࣭ศἪࡢㄪ⠇ࠊ(4)㦵 ⱆ-◚㦵⣽⬊ࢿࢵࢺ࣮࣡ࢡࠊ࡞ࠊ」㞧࡞㐣⛬ࡀᶵ⬟ ⓗ⤖ࡧࡘ࠸࡚⦓ᐦ࡞ไᚚࡀ⾜ࢃࢀ࡚࠸ࡿ(ᅗ 1)ࠋᐇ 㝿ศࡢྛẁ㝵ࡢᚲせᛶᛂࡌ࡚ࠊNFATc1 ࡀࠊ TRAP (tartrate-resistance acid phosphatase), FDOFLWRQLQ UHFHSWRU FDWKHSVLQ . ȕ- integrin ࡞ࡢ ⏕⌮స⏝ࡢ␗࡞ࡿከᙬ࡞◚㦵⣽⬊≉␗ⓗ㑇ఏᏊࢆ ḟࠎ㌿άᛶࡍࡿࡀࠊࡇࢀࡽࡢከ✀ከᵝ࡞ୗὶ 㑇ఏᏊ⩌ࢆࡢࡼ࠺࡞ศᏊ࣓࢝ࢽࢬ࣒࡛ࠊ✵㛫ⓗ࣭ 㛫ⓗ࡞࠸ศࡅࢆࡋ࡚࠸ࡿࡀ༑ศゎ᫂ࡉࢀ࡚࠸ ࡞࠸ࠋ (2)◊✲ࡢ┠ⓗ ᡃࠎࡣࡇࢀࡲ࡛ࠊヨ⸆ῧຍᚋࡢ㦵ⱆ⣽⬊ࡽ᥇ ྲྀࡋࡓ conditioned medium ࢆ⏝࠸࡚ࠊࡑࡢ◚㦵⣽⬊ ศ࠼ࡿᙳ㡪ࢆ᳨ウࡍࡿᐇ㦂⣔ࢆ☜❧ࡋࠊ RAGE ࢩࢢࢼࣝࡼࡿ◚㦵⣽⬊ศ࠾ࡅࡿ㦵ⱆ⣽ ⬊ࡽࡢ paracrine signaling ࡢ㔜せᛶࢆ᫂ࡽࡋ ࡓࠋࡲࡓࠊ㦵ⱆ⣽⬊-◚㦵⣽⬊ࡢࢡࣟࢫࢺ࣮ࢡゎᯒࢆ ࡍࡿࡓࡵࡢࠊ㦵ⱆ⣽⬊㸭◚㦵⣽⬊ඹᇵ㣴ᐇ㦂⣔ࢆᵓ ⠏ࡋ࡚࠸ࡿࠋࡲࡓࠊ㌿ᅉᏊ」ྜయࡼࡿࢡ࣐ࣟࢳ ࣭࢚ࣥࣆࢪ࢙ࢿࢸࢡࢫࡢᶵ⬟ไᚚࢆࢤࣀ࣒࣡ࢻ ࡛ゎᯒࡍࡿᡭἲࢆ㛤Ⓨࡋ࡚࠸ࡿࠋᮏ◊✲࡛ࡣࠊࡇࢀ ࡽࡢࢩࢫࢸ࣒ࢆᛂ⏝࣭⼥ྜࡋࠊ◚㦵⣽⬊ศࡢ࣐ࢫ ࢱ࣮㌿ᅉᏊ NFATc1 ࢆゎᯒᑐ㇟ࡋ࡚ࠊ⏕Ꮫⓗ ࡞㌿」ྜయゎᯒᢏ⾡ḟୡ௦ᆺࢩ࣮ࢡ࢚ࣥࢧ࣮ ࡼࡿࢤࣀ࣒࣡ࢻࡢ࢚ࣆࢪ࢙ࢿࢸࢡࢫ࣭ࢺࣛࣥࢫ ࢡࣜࣉࢺ࣮࣒ゎᯒࢆ᭷ᶵⓗ⤖ࡧࡘࡅࡿࡇ࡛ࠊ㦵 ࣜࣔࢹࣜࣥࢢไᚚࡢ᪂ࡓ࡞ศᏊᶵᵓࣉ࣮ࣟࢳࡋࠊ 㦵࢚ࣆࢪ࢙ࢿࢸࢡࢫࡢศᏊែゎ᫂⸆ᇶ┙ࡢ 㛤Ⓨࢆヨࡳࡿࡇࢆ┠ⓗࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 RNA-sequence 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 - 418 - ◚㦵⣽⬊ศ࠾࠸࡚ࡣࠊ◚㦵⣽⬊๓㥑⣽⬊ coding genesࡢⓎ⌧ቑຍࡀㄆࡵࡽࢀࡓ㸦ᅗ㸰㸧ࠋࡑ (Monocyte/ Macrophage⣔⣽⬊)ࡀ⣽⬊ቑṪ࣭ศ࣭ ࡢഴྥࡣ㸵᪥ᚋࡲ࡛⥆ࡁࠊ㸯㸲᪥ᚋࡣ㑇ఏᏊⓎ⌧ άᛶࢆ⤒࡚ᙧែⓗࡶᶵ⬟ⓗࡶⓗ࡞ኚࢆ㐙 ࡣprotein-coding, non-coding genesࡶⓗῶᑡ ࡆࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋ≉◚㦵⣽⬊ࡢᡂ⇍࠾ ࡋࡓࠋ ࠸࡚ࡣDcstampࡸOcstamp࡞ࡢ⺮ⓑࢆࡋࡓ◚㦵 ⣽⬊ࡢ⼥ྜ࣭ከゅᕧ⣽⬊ࡸSealing zoneᅖࡲࢀࡓ 㦵྾❐ࡢᙧᡂࠊἼ≧⦕࠾ࡅࡿ࢝ࢸࣉࢩࣥKࡸ࣐ࢺ ࣜࢵࢡࢫ࣓ࢱࣟࣉࣟࢸ࣮ࢮࡢⓎ⌧ࠊࣉࣟࢺ࣏ࣥࣥ ࣉࡼࡿ㦵྾❐ࡢ㓟ᛶࡀ㔜せ࡛࠶ࡾࠊ◚㦵⣽⬊ ศ㐣⛬࠸࡚ከࡃࡢᶵ⬟ⓗ␗࡞ࡿ㑇ఏᏊࡢⓎ ⌧ࡀ㛫ⓗ࣭✵㛫ⓗᕦጁㄪ⠇ࡉࢀ࡚࠸ࡿࡇࡀ ணࡉࢀࡿࠋ◚㦵⣽⬊ศ࠾࠸࡚ࡣ㦵ⱆ⣽⬊ࡸࢫ ࢺ࣮࣐ࣟ⣽⬊࡞ࡽศἪࡉࢀࡿReceptor activator of NF-ț% OLJDQG 5$1./ 0DFURSKDJH FRORQ\ stimulating factor (M-CSF)࡞ࡢࢧࢺ่⃭࢝ࣥ ࡀ㔜せ࡛࠶ࡿࡀࠊࡇࢀࡽࡢ่⃭ࡼࡾࡢࡼ࠺࡞㑇 ఏᏊࡀࡢ㛫ᖏⓎ⌧ㄏᑟࡉࢀ࡚࠸ࡿࡢࡘ࠸ ࡚ࡣࠊᮍࡔ᫂ࡽ࡛࡞࠸㒊ศࡀከ࠸ࠋࡑࡇ࡛ᡃࠎࡣࠊ ๓◚㦵⣽⬊⣔⣽⬊ RAW264.7⣽⬊࠾ࡅࡿRANKL ౫Ꮡᛶቑຍࡍࡿ㑇ఏᏊࡘ࠸࡚RNA-seqࢆ⏝࠸࡚ ᳨ウࢆ⾜ࡗࡓ㸦ᅗ1㸧ࠋMicro-array␗࡞ࡾࠊRNA-seq ࡉࡽࠊࡑࢀࡒࢀࡢ㛫࠾ࡅࡿⓎ⌧ቑຍ⋡ࡢ㧗 ࠸㑇ఏᏊࡘ࠸࡚ୖ30㑇ఏᏊࢆ☜ㄆࡋࡓࡇࢁࠊ ่⃭ᚋ㸰᪥┠ࡣ᰾ෆࢩࢢࢼࣝ㛵ࡍࡿ⒴㑇ఏᏊ MycࡸTNF receptorࢩࢢࢼࣝࡢ⣽⬊ෆࡢఏ㐩㔜 せ࡞TRAF1ࠊࡲࡓsmall nucleolar RNA, small nuclear RNA, micro RNA, transfer RNA ࡞ ከ ࡃ ࡢ non-coding RNAࡀྠᐃࡉࢀࡓࠋ୍᪉ࠊ่⃭ᚋ㸱᪥┠ ࡞ࡿ◚㦵⣽⬊ศࡢmaster regulator࡛࠶ࡿ㌿ ᅉᏊNuclear factor of activated T-cells, cytoplasmic 1 (Nfatc1)ࡢⓎ⌧ቑຍ㐃ືࡋ࡚Mmp9, Nhedc2, Ctsk, Atp6v0d2࡞◚㦵⣽⬊ࡢ㦵྾ᶵ⬟㔜せ࡞㑇ఏᏊ ࡢⓎ⌧ቑຍࢆྵࡵࠊୖ༨ࡵࡿ㑇ఏᏊࡢṤࡀ protein-coding genes࡛࠶ࡗࡓ㸦⾲㸯㸧ࠋ㸵᪥┠ࡣ ࡛ࡣnon-coding RNAࢆྵࡵࡓゎᯒࢆ⾜࠺ࡓࡵࠊࡼࡾ ⥙⨶ⓗ㑇ఏᏊኚࢆᤊ࠼ࡿࡇࡀྍ⬟࡛࠶ࡾࠊᐇ 㝿ᅗ1♧ࡍࡼ࠺࡞Micro-RNA (Mir1902, Mir762, Mir2861) ࢆࡣࡌࡵࡋ࡚ᵝࠎ࡞non-coding RNAࡀ RANKL౫ᏑⓗⓎ⌧ኚࢆ᮶ࡋ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋ≉⯆῝࠸Ⅼࡋ࡚ࠊྠࡌRANKLୗ ὶ㑇ఏᏊ࡛ࡶⓎ⌧ࡢቑຍࡍࡿᮇࡀ␗࡞ࡾࠊRANKL ่ ⃭ ᚋ ᪩ ᮇ ࡣ signal ఏ 㐩 㛵 ࡍ ࡿ 㑇 ఏ Ꮚ ࡸ ୖ30㑇ఏᏊࡢ࡚ࡀprotein-coding genes࡛࠶ࡾࠊ non-coding RNAࡀቑຍࡋ࡚࠸ࡿࡇࡀ᫂ࡽ ◚㦵⣽⬊⼥ྜ࣭ከ᰾ᕧ⣽⬊㔜せ࡞Ocstampࡸ◚ ࡞ࡗࡓ୍᪉ࠊRANKL่⃭3᪥ᚋࡽࡣከࡃࡢprotein 㦵⣽⬊άᛶ㔜せ࡞Acp5࡞ࡢⓎ⌧ࡀ㧗࠸ࡇࡀ - 419 - ࡀ࠶ࡿ୍᪉࡛ࠊOscar, Clcn7, Serpind1࡞ࡢ㸰┦ᛶ ࡢⓎ⌧ኚࢆ♧ࡍ㑇ఏᏊࡸࠊCar2, Acp5࡞ࡢⓎ⌧ ࡢࣆ࣮ࢡࡀNfatc1␗࡞ࡿ㑇ఏᏊࡀ☜ㄆࡉࢀࡓ㸦ᅗ 㸱㸧ࠋࡇࡢࡇࡣࠊࡇࢀࡽࡢ㑇ఏᏊࡢⓎ⌧Nfatc1 ࡀ㛵ࡍࡿࡶࡢࡢࠊNfatc1௨እࡢࢩࢢࢼࣝࡽࡢ㑇 ఏᏊⓎ⌧ㄪ⠇ࡸNFATc1ࡢ㌿ᚋಟ㣭ࡢᏑᅾࢆ♧၀ ࡋ࡚࠾ࡾࠊ௨ୖࡢ⤖ᯝࡽࠊ◚㦵⣽⬊ศ࠾ࡅࡿ ከᙬ࡞㑇ఏᏊⓎ⌧ㄪ⠇ᶵᵓࡢᏑᅾࡀ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢤࣀ࣒ᨭࡼࡗ࡚ᚓࡽࢀࡓゎᯒ⤖ᯝࡽࠊ ☜ㄆࡉࢀࠊRANKL่⃭ᚋ㸵᪥┠ࡢRAW264.7⣽⬊ࡀ RANKL౫Ꮡᛶ◚㦵⣽⬊ศ࠾ࡅࡿdynamic࡞㑇ఏ ◚㦵⣽⬊ࡋ࡚ᡂ⇍ẁ㝵࠶ࡿࡇࡀ᥎ ࡉࢀࡓࠋ ᏊⓎ⌧ㄪ⠇ᶵᵓࡼࡾࠊ◚㦵⣽⬊ࡢከᙬ࡞ᶵ⬟࣭ᙧ ࡇࢀࡲ࡛ࡢ᳨ウࡽࠊRANKLୗὶ㑇ఏᏊࡑࢀࡒࢀ ែኚࡀࡶࡓࡽࡉࢀ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋࡲ ࠾࠸࡚Ⓨ⌧ቑຍࡍࡿᮇࡀ␗࡞ࡿࡇࡀ☜ㄆࡉࢀ ࡓࠊ㌿ᅉᏊNFATc1ࡢ⩻ヂᚋಟ㣭ࡢᏑᅾࡶ♧၀ࡉࢀ ࡓࡀࠊ㌿ᅉᏊNFATc1ࡢୗὶ㑇ఏᏊࡶྠᵝࡢ㇟ ࡓࡓࡵࠊ⌧ᅾᡃࠎࡣTandem affinity purification ࡀ ㄆ ࡵ ࡽ ࢀ ࡿ ࡘ ࠸ ࡚ ࠊ 㐣 ཤ conditional LC-MS/MSࡼࡾྠᐃࡋࡓ᪂ࡓ࡞NFATc1⤖ྜศᏊ knockout ࡢᶵ⬟ゎᯒࢆ⾜ࡗ࡚࠾ࡾࠊࡇࢀࡽࡢ⤖ᯝࡘ࠸࡚⌧ mice ࢆ ⏝ ࠸ ࡚ ྠ ᐃ ࡉ ࢀ ࡓ NFATc1-dependent genes (Aliprantis AO, et al. J ᅾࠊㄽᩥᢞ✏‽ഛ୰࡛࠶ࡿࠋ Clin Invest 2008; Charles JF, et al. Bone 2012) ࢆཧ ⪃᳨ウࡋࡓࠋࡍࡿணࡋࡓ㏻ࡾࠊCalcr, Ctsk, Mmp9࡞Nfatc1㢮ఝࡢⓎ⌧ኚࢆㄆࡵࡿ㑇ఏᏊ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᪥ᮏෆศἪᏛࠊ࣓ࣜ࢝ෆศἪᏛ - 420 - බເᨭㄢ㢟㸸2010 ᖺᗘ 2012 ᖺᗘ S ㌿ᅉᏊ」ྜయࡼࡿࢡ࣐ࣟࢳࣥᶵ⬟ㄪ⠇ L36 ࣜࣉࣟࢢ࣒ࣛไᚚᶵᵓࡢゎ᫂ ᇶ┙◊✲㸦㹀㸧 ⏣୰ ▱࣭᫂༓ⴥᏛᏛ㝔་Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 ㏆ᖺࡢ⏕་⒪ࡢ㐍Ṍࡣࠊ┠ぬࡲࡋ࠸㐍Ṍࢆ㐙ࡆ࡚ ࠾ࡾࠊ⮫ᗋᛂ⏝ࡢᮇᚅࡀ㧗ࡲࡗ࡚࠸ࡿࠋࡋࡋࡑ ࡢ୍᪉࡛ࠊᖿ⣽⬊⥔ᣢࡸศไᚚࡘ࠸࡚ࡢヲ⣽࡞ ࣓࢝ࢽࢬ࣒ࡣᮍ▱ࡢⅬࡶከ࠸ࠋࡑࡢ୰࡛ࠊᡃࠎࡣᖿ ⣽⬊ไᚚศࡢ࣮࢟ࣇࢡࢱ࣮ࡋ࡚㌿ᅉᏊ p53 ╔┠ࡋࡓࠋp53 ࡣ⒴ᢚไ㑇ఏᏊࡋ࡚ྂࡃࡽ ▱ࡽࢀ࡚࠸ࡿࡀࠊ㏆ᖺ᰾ࣜࣉࣟࢢ࣑ࣛࣥࢢࡢไᚚࡸ ES ⣽⬊ࡢึᮇศࡢ㛵ࡀ᫂ࡽ࡞ࡗ࡚࠸ࡿࠋ ᮏ◊✲࡛ࡣࠊS ࡸ᰾ࡢึᮇᚲ㡲ࡢ㌿ᅉᏊࢆ୰ ᚰࠊࢡ࣐ࣟࢳࣥᶵ⬟࢚ࣆࢪ࢙ࢿࢸࢡࢫࢆไᚚ ࡍࡿ㌿ᅉᏊ」ྜయࢿࢵࢺ࣮࣡ࢡ㔜せ࡞ศᏊࢆ⥙ ⨶ⓗྠᐃࡍࡿࡇ࡛ࠊᖿ⣽⬊ไᚚࡢ᰿ᗏ࡛స⏝ࡍ ࡿ᰾ෆࢩࢢࢼࣝࢡ࣐ࣟࢳࣥไᚚ࣓࢝ࢽࢬ࣒ศᏊ ⏕Ꮫⓗࣉ࣮ࣟࢳࡋࠊḟୡ௦ᆺࢩ࣮ࢡ࢚ࣥࢧ࣮ ࡼࡿࢤࣀ࣒࣡ࢻࡢ HSLJHQHWLFV࣭ࢺࣛࣥࢫࢡࣜࣉ ࢺ࣮࣒ゎᯒ᭷ᶵⓗ⤖ࡧࡘࡅࡿࡇ࡛ࠊຠ⋡ⓗ࡛ 㠉᪂ⓗ࡞ L36 స〇ࠊศไᚚᑐࡍࡿᢏ⾡ࡸࡘ࡞ ࡆࡿࠋ 㸺ᨭࡢෆᐜ㸼 ḟୡ௦ᆺࢩ࣮ࢡ࢚ࣥࢧ࣮ࡼࡿ QRQFRGLQJ51$ ࢆྵ ࡵࡓ㌿⏘≀ࡢ࢚ࣆࢪ࢙ࢿࢸࢡࢫ࣭ࢺࣛࣥࢫࢡࣜ ࣉࢺ࣮࣒ไᚚࡢゎᯒࡘ࠸࡚ᨭࢆཷࡅࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ES/iPS ⣽⬊࠾ࡅࡿࢤࣀ࣒࣡ࢻࡢゎᯒࢆ⾜࠺ࡇ ࡼࡾࠊ(6L36 ≉␗ⓗⓎ⌧ࡍࡿ S ౫Ꮡⓗ࡞ QRQFRGLQJ51$ ࢆྵࡵࡓ㌿⏘≀ࡢ࢚ࣆࢪ࢙ࢿࢸ ࢡࢫ࣭ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ไᚚࡢゎᯒࢆ⾜ࡗࡓࠋ ࡑࡢ୰࡛ࡶ≉ࠊL36 (6 ඹ㏻ࡍࡿ㛗㙐ࡢ⺮ⓑ ⩻ヂࡉࢀ࡞࠸ 51$LQF51$ೃ⿵ࡀ 㑇ఏᏊ᳨ฟࡉ ࢀࡓࠋ ≉ &KU ࡽㄞࡳฟࡉࢀࡿ OLQF51$ 㛵ࡋ࡚ࠊ &K,3VHT ࢆྜࢃࡏ࡚⾜ࡋ࡚ࠊࡑࡢⓎ⌧ࣉࣟࣇ ࣝゎᯒ HSLJHQHWLF ࣃࢱ࣮ࣥࢆゎᯒࡋࡓࠋ766 ゎᯒ ࡚㌿㛤ጞⅬࡢ᫂ࡽ࡞ࣆ࣮ࢡࢆㄆࡵࠊ51$VHT ࡽ⣙ NE Ώࡿ㌿⏘≀ࡀ☜ㄆࡉࢀࡓࠋ㌿ 㛤ጞⅬ㡿ᇦ㏆ഐ 1$12* ࡢ⤖ྜࡀほᐹࡉࢀࠊࡲࡓㄞ ࡳฟࡋ㡿ᇦ୍⮴ࡋ࡚ࠊDFWLYH ࣄࢫࢺࣥࢥ࣮ࢻ࡛࠶ ࡿ $F.+ࠊPH.+ ྠࠊUHSUHVVLYH ࣄࢫࢺ ࣥࢥ࣮ࢻ࡛࠶ࡿ PH.+ ࡢࣆ࣮ࢡࡀㄆࡵࡽࢀࡓࠋ ࡇࢀࡽࡢ⤖ᯝࡣࠊELYDOHQW ࡞ HSLJHQHWLFUHJXODWLRQ ࢆཷࡅࡿ OLQF51$ ࡢᏑᅾࢆ♧၀ࡋ࡚࠸ࡓࠋࡉࡽࠊ S ౫Ꮡⓗ '1$ യᐖ࡚㌿ㄏᑟࢆཷࡅࡿ OLQF51$ ೃ⿵ࢆ 㑇ఏᏊྠᐃࡋࡓࠋࡇࢀࡽࡢ OLQF51$ ᑐ ࡋ࡚ࠊVK51$ ࡼࡿᶵ⬟ⓗࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡋ ࡓᡤࠊ୍㒊ࡢ OLQF51$ ࡛ (6L36 ࡢ SOXULSRWHQF\ ࡀ ኻࢃࢀࠊ(6L36 ࡢከ⬟ᛶไᚚࡢ୍㒊ࡢ࣓࢝ࢽࢬ࣒ ࡋ࡚ࡇࢀࡽࡢ OLQF51$ ࡀ㛵ࡋ࡚࠸ࡿࡢᛶࡀ♧၀ ࡉࢀࡓࠋ ࡉࡽᡃࠎࡣࠊ ࣄࢺ ES/iPS ⣽⬊ศไᚚ࠾ࡅࡿ p53 ࡢⓎ⌧࣭ᶵ⬟ゎᯒࢆ⾜ࡗࡓࠋDual SMAD inhibition ࡢ ⣔ࢆ⏝࠸࡚⚄⤒ᖿ⣽⬊ࡢศࢆ⾜࠺ࠊ⚄⤒ᖿ⣽ ⬊ࡢ࣐ࢫࢱ࣮ࣞࢠ࣮ࣗࣞࢱ࣮࡛࠶ࡿ PAX6 ࡢⓎ⌧ୖ ᪼ࢆㄆࡵࡿࡀࠊࡇࡢศࡢ㐣⛬࠾࠸࡚ࠊp53 ࡀ┦ ᛶㄏᑟࡉࢀࡿࡇࡀほᐹࡉࢀࡓࠋࡑࡇ࡛ࠊࡇࢀࡽ ࡢ࣓࢝ࢽࢬ࣒ࢆ᫂ࡽࡍࡿᙜࡓࡾࠊCRISPR/Cas9 ࢩࢫࢸ࣒ࢆ⏝࠸࡚ p53 ࣀࢵࢡ࢘ࢺࣄࢺ ES/iPS ⣽⬊ ࢆసᡂࡋゎᯒࢆ⾜ࡗࡓࠋ⚄⤒ᖿ⣽⬊ศ࠾ࡅࡿ p53 ࣀࢵࢡ࢘ࢺࣄࢺ ES/iPS ⣽⬊ࡢゎᯒ࡛ࡣࠊPAX6 ࡢ Ⓨ⌧ࡀ㔝⏕ᆺẚࡋ࡚ࡼࡾ᪩ᮇࡽㄏᑟࡉࢀࡿࡇ ࡀ♧ࡉࢀࠊp53 ࣀࢵࢡ࢘ࢺࣄࢺ ES/iPS ⣽⬊࡛ࡣ single SMAD inhibition ࡛ࡶ PAX6 ࡀᙉࡃㄏᑟࡉࢀࡿ ࡇࡀ♧ࡉࢀࡓࠋࡇࢀࡽࡼࡾ p53 ࡣ ES/iPS ⣽⬊ࡢ⚄ ⤒ᖿ⣽⬊ࡢศࡢ㐣⛬࡛ PAX6 ࢆ㈇ไᚚࡋ࡚࠸ ࡿ⪃࠼ࡽࢀࡓࠋࡇࡢ࣓࢝ࢽࢬ࣒ゎᯒ࡛ࡣࠊࢡ࣐ࣟ ࢳࣥචỿ㝆ἲࢆ⏝࠸࡚ࠊPAX6 ࡢ࢚ࣆࢪ࢙ࢿࢸࢵ ࢡಟ㣭≧ែࢆㄪࡓࡇࢁࠊp53 ࣀࢵࢡ࢘ࢺ⣽⬊࡛ ࡣ⚄⤒ᖿ⣽⬊ࡢศ㐣⛬࡛ H3K4 ࣓ࢳࣝࡀ㔝⏕ᆺ ẚࡋ࡚ᙉࡃㄏᑟࡉࢀࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡉ ࡽࠊSCID ࣐࢘ࢫࡢ⢭ᕢୖ⬡⫫ ES/iPS ⣽⬊ࢆᡴ ࡕ㎸ࡳࠊࢸࣛࢺ࣮࣐ᙧᡂࢆほᐹࡋࡓᡤࠊp53 ࣀࢵࢡ ࢘ࢺ⣽⬊࡛ࡣ⬻⤌⧊ࡀࡼࡾከࡃ࡞ࡿഴྥࢆㄆࡵࡓࠋ ࡇࢀࡽࡢ⤖ᯝࡣࠊp53 ࡀࣄࢺ ES ⣽⬊ࡢ⚄⤒ᖿ⣽⬊ ࡢึᮇศ࠾࠸࡚ PAX6 ࡢ repressive ࡞Ⓨ⌧ㄪ⠇ࢆ ࡋ࡚ࠊ⚄⤒ᖿ⣽⬊ศࢆᢚไࡋ࡚࠸ࡿྍ⬟ᛶࢆ♧ ࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢤࣀ࣒ᨭࡼࡿࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ㏻ࡌ ࡚ࠊES/iPS ⣽⬊࠾ࡅࡿ p53 ౫Ꮡⓗస⏝ࡍࡿ linc RNA ࡢᏑᅾࠊࡑࢀࡽࡀከ⬟ᛶ⥔ᣢ㛵ࡋ࡚࠸ࡿྍ ⬟ᛶࡀ᫂ࡽ࡞ࡗࡓࠋࡉࡽࠊp53 ࡀࣄࢺ ES/iPS ࡢ୕⬇ⴥ⣔ࡢึᮇศ࠾࠸࡚ࠊ୍㒊ࡢศ㛵 ࡍࡿ㑇ఏᏊࡢ࢚ࣆࢪ࢙ࢿࢸࢵࢡไᚚࢆኚࡉࡏࠊ ࡇࢀࡽࡢไᚚࢆ⾜ࡗ࡚࠾ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ ࡇࢀࡽࡢᡂᯝࡼࡾ p53 ࢆไᚚࡍࡿࡇࡼࡾࠊ ES/iPS ⣽⬊ࡢࣜࣉࣟࢢ࣑ࣛࣥࢢࡸศไᚚࢆຠ⋡ ࡼࡃ⾜࠺ࡇࡀྍ⬟ᛶ࡞ࡿ࠸࠺ᒎᮃࡀ㛤ࡅࡓ ⪃࠼ࡽࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Suzuki, S. et al., PNAS 107, 7461-7466 (2010) Horiuchi, S. et al., J Immunol 186, 6378-6389 (2011) Hosokawa, H. et al., PNAS 110, 4691-4696 (2013) Hosokawa, H. et al., PNAS 110, 18626-18631 (2013) - 421 - බເᨭㄢ㢟㸸2010 ᖺᗘ ㌿ᅉᏊ p53 ࡼࡿ᪂ࡓ࡞௦ㅰㄪ⠇ᶵ⬟௦ㅰ⎔ቃᛂ⟅ࡢ࢚ࣆࢪ࢙ࢿࢸࢡࢫไᚚ ᣮᡓⓗⴌⱆ◊✲ ⏣୰ ▱࣭᫂༓ⴥᏛᏛ㝔་Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 ⢾ᒀࡸ⫧‶࡞ࡢ⏕ά⩦័ࡣᵝࠎ࡞⒴ࡢⓎ ࢆቑຍࡉࡏࡿࡇࡀ᫂ࡽࡉࢀ࡚࠸ࡿࠋ୍᪉࡛ࠊ ⒴ᢚไ㑇ఏᏊࡶ⣽⬊࿘ᮇṆ࣭࣏ࢺ࣮ࢩࢫ࣭⪁ ࢆࡋࡓ⒴ᢚไᶵ⬟ࡢࡳ࡛࡞ࡃ࣮࢜ࢺࣇࢪ࣮ࠊ࢚ ࢿࣝࢠ࣮௦ㅰࠊᢠ㓟స⏝࡞ከᒱΏࡾ⣽⬊ෆ௦ ㅰไᚚࢆไᚚࡋ࡚࠸ࡿࡇࡀḟࠎ᫂ࡽ࡞ࡗ࡚ ࠾ࡾ⒴ࡢᯟ⤌ࡳࢆ㉸࠼࡚⏕≀Ꮫⓗࣉࣟࢭࢫ㔜せ࡛ ࠶ࡿ⪃࠼ࡽࢀ࡚࠸ࡿࠋࡑࡋ࡚⒴⏕ά⩦័ࢆ⤖ ࡧࡅࡿศᏊែࡢ᰿ᗏࠊp53 ࡼࡿ࢚ࢿࣝࢠ࣮࣭ ௦ㅰㄪ⠇ᶵᵓࡢᏑᅾࡀ᥎ᐃࡉࢀ࡚࠸ࡿࠋࡋࡋ࡞ࡀ ࡽࠊᐇ㝿 p53 ࡀࡢࡼ࠺࡞࣓࢝ࢽࢬ࣒ࢆࡋ࡚⏕ ά⩦័ࡢែᙧᡂࠊ᭦ࡣ⏕ά⩦័ྜేࡍࡿ ⒴ࡢⓎ㛵ࡋ࡚࠸ࡿࠊࡑࡢᐇయࡣ༑ศゎ᫂ ࡉࢀ࡚࠸࡞࠸ࠋ 㸺ᨭࡢෆᐜ㸼 ᡃࠎࡣࢤࣀ࣒ᨭࡼࡾḟୡ௦ᆺࢩ࣮ࢡ࢚ࣥࢧ࣮ ࢆ⏝࠸ࡓ ChIP-seq ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ ࡼࡿ⣽⬊ෆ௦ㅰࢆไᚚࡍࡿ p53 ୗὶ㑇ఏᏊ ࡋ࡚ ࢢࣝࢱ࣑ࣥ௦ㅰࢆྖࡿ Glutaminase2(GLS2)ࡸ᰾㓟 ௦ㅰ㛵ࢃࡿ㓝⣲㧗࠸┦ྠᛶࢆ᭷ࡍࡿ DPYSL4 ࡞ ⯆῝࠸㑇ఏᏊࢆ࠸ࡃࡘྠᐃࡋ࡚ࡁࡓࠋGLS2 ࡣ࣑ࢺࢥࣥࢻࣜᒁᅾࡋࢢࣝࢱ࣑ࣥࢆຍỈศ ゎࡋࢢࣝࢱ࣑ࣥ㓟ࢆ౪⤥ࡍࡿ⣽⬊ෆ௦ㅰ㓝⣲࡛࠶ࡿࠋ ᭦࣓ࢱ࣮࣒࣎ࣟゎᯒࡸ⏕Ꮫⓗᡭἲ⤌ࡳྜࢃࡏ ࡿࡇࡼࡾ GLS2 ࡣࢢࣝࢱ࣑ࣥ㓟ࡽȘࢣࢺࢢࣝ ࢱࣝ㓟ࢆࡋ࡚ TCA ᅇ㊰స⏝ࡋዲẼⓗᅇ㊰ࢆ⤒ ࡚ ATP ⏘⏕㓟⣲ᾘ㈝㔞ࢆṇྖࡿ⤌ࡳࢆ᫂ࡽ ࡋሗ࿌ࡋ࡚ࡁࡓࠋࡑࡇ࡛⏕ά⩦័⒴ࡢ᥋Ⅼ ࡋ࡚ NASH ╔┠ࡋࠊp53-GLS2 ࡀࡇࢀࡽ⣽⬊ෆ௦ ㅰไᚚࢆࡋ࡚⏕ά⩦័⏕ά⩦័క࠺ NASH-⫢⒴Ⓨ㛵ࡍࡿࡢ GLS2 ࣀࢵࢡ࢘ࢺ ࣐࢘ࢫࢆసᡂࡋ⏕యෆ࠾ࡅࡿᙺࢆヲ⣽᳨ウࡋ ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 GLS2 ࡣ In vitro ࠾࠸࡚ p53 ୗὶ㑇ఏᏊࡋ࡚ ⒴ࡢ⣽⬊ቑṪᢚไࠊmigration, invasion ᢚไస⏝ࢆ ᭷ࡋ࡚࠾ࡾࠊGLS2 ࢆࣞࢺࣟ࢘ࣝࢫࢆ⏝࠸࡚㐣 Ⓨ⌧ࡉࡏࡓ⫵⒴⣽⬊ࢆ SCID ࣐࢘ࢫࡢ⓶ୗ⛣᳜ࡋ ࡚ࡶ⒴ᢚไຠᯝࢆ♧ࡍࡇࡀ♧ࡉࢀࡓࠋ GLS2 ࡣ࣐࢘ࢫ࠾࠸࡚⫢⮚ࠊ⬻ࠊ⮅⮚࠸ࡗࡓ ⣽⬊ෆ௦ㅰ῝ࡃ㛵ࢃࡗ࡚࠸ࡿ⮚ჾ㧗Ⓨ⌧ࡋ࡚࠸ ࡓࠋ᭦ࠊ㧗⬡⫫㣗㈇Ⲵࡋࡓ࣐࢘ࢫ⫧‶ࣔࢹࣝ࠾ ࠸࡚࣐࢘ࢫࡢ⬡⫫ࠊ⬡⫫⫢ࠊNASH ⫢࡛ GLS2 ࡢⓎ ⌧ࡀ᭷ពቑຍࡋ࡚࠾ࡾఱࡽࡢᙺࢆᯝࡓࡋ࡚࠸ ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ ࡑࡇ࡛ GLS2 ࣀࢵࢡ࢘ࢺ࣐࢘ࢫࢆసᡂࡋ㧗⬡⫫ 㣗ࢆ㈇Ⲵࡋ GLS2 ࡢ⏕యෆ࠾ࡅࡿᙺࢆ᳨ウࡋࡓࠋ ࡲࡎ GLS2 ࣀࢵࢡ࢘ࢺ࣐࢘ࢫࡣ࣓ࣥࢹࣝࡢἲ๎ ᚑࡗ࡚⏕ࡲࢀࡿࡇࡀ☜ㄆࡉࢀࡓࠋ᭦⯆῝࠸ࡇ ࠊ㧗⬡⫫㣗㈇Ⲵ GLS ࣀࢵࢡ࢘ࢺ࣐࢘ࢫࡣ㧗⬡ ⫫㣗㈇Ⲵ WT ࣐࢘ࢫẚ㍑ࡋ࡚᫂ࡽ⫧‶ࠊ⪏⢾ ⬟㞀ᐖࠊ㧗୰ᛶ⬡⫫⾑࠸ࡗࡓ࠸ࢃࡺࡿ࣓ࢱ࣎ࣜ ࢵࢡೃ⩌ࢆ࿊ࡋ࡚࠸ࡓࠋ㧗⬡⫫㣗 GLS2 ࣀࢵࢡ ࢘ࢺ࣐࢘ࢫࡣ⬡⫫⫢ࡸ NASH ࢆ࿊ࡋࡓࡀࠊ⫢⒴ࡢⓎ ࡣㄆࡵࡽࢀ࡞ࡗࡓࠋࡑࡇ࡛ࢫࢺࣞࣉࢺࢰࢻࢩࣥ ⓶ୗὀᑕ㧗⬡⫫㣗㈇Ⲵࢆ⤌ࡳྜࢃࡏ࡚ NASH-⫢ ⒴Ⓨ࣐࢘ࢫࣔࢹ࡛ࣝ࠶ࡿ STAM ࣐࢘ࢫࢆసᡂࡋࠊ ⫢⒴࠾ࡅࡿᙺࢆ᳨ウࡋࡓࠋࡑࡢ⤖ᯝ STAM Ⓨ GLS2 ࣀࢵࢡ࢘ࢺ࣐࢘ࢫࡣ WT ẚ㍑ࡋ࡚ࡼࡾ᪩ ᮇ⫢⒴ࢆⓎࡍࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋNAH⫢⒴Ⓨᡃࠎࡣ GLS2 ࡢᢠ㓟స⏝ࡀ㔜せ࡞ᙺ ࢆᯝࡓࡋ࡚࠸ࡿࡢ࡛ࡣ࡞࠸᥎ᐃࡋࠊ⌧ᅾᢠ㓟 NAC ࢆ⏝࠸࡚᭦᳨ウࢆࡍࡍࡵ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ⏕యෆ࠾࠸࡚ p53 ࡼࡗ࡚ไᚚࡉࢀࡿࢢࣝࢱ࣑ࣥ ࡢ࣐ࢫࢱ࣮ࣞࢠ࣮ࣗࣞࢱ࣮࡛࠶ࡿ GLS2 ࡣ⏕ά⩦័ ࡢⓎࠊ᭦ࡣ⬡⫫⫢-NASH-⫢⒴Ⓨ῝ࡃ㛵 ࡍࡿࡇࢆぢฟࡋࡓࠋNASH ࡣ⏕ά⩦័ࡢቑຍ క࠸Ⓨᩘࡣቑຍࡋ࡚࠸ࡿࡀࠊNASH ࡢⓎᶵᗎࡣ ᫂࡞Ⅼࡀከࡃࠊ☜ᐃデ᩿ࡣくⓗ࡞⫢⏕᳨ࡀᚲ せ࡛࠶ࡿࠋ᭦ NASH ࡣ⫢◳ኚ࣭⫢⒴ࡢ㐍⾜ࢆ㜵 ࡄࡓࡵ⏕ά⩦័ࡢᨵၿຍ࠼࡚ࠊࣥࢫࣜࣥᢠ ᛶᨵၿ⸆ࣆ࢜ࢢࣜࢲࢰࣥࡸࣅࢱ࣑ࣥ E ࡞ࡢ⸆≀ ⒪ࡀ⾜ࢃࢀ࡚࠸ࡿࡀࠊ⌧≧࡛ࡣ NASH ᑐࡍࡿ᭷ ຠ࡞⒪ἲࡀ࡞࠸ࡇࡀࡁ࡞ၥ㢟࡞ࡗ࡚࠸ࡿࠋ ࡼࡗ࡚ GLS2 ࡢ NASH-⫢⒴Ⓨ࠾ࡅࡿヲ⣽࡞ศ Ꮚែࡢゎ᫂ࡣ᪂ࡋ࠸ࢱࣉࡢ⒪㛤Ⓨ⤖ࡧࡃ ྍ⬟ᛶࢆ⛎ࡵ࡚࠸ࡿࠋࡲࡓ㏆ᖺࢢࣝࢱ࣑ࣥ௦ㅰࡸȘ ࢣࢺࢢࣝࢱࣝ㓟࡞ࡢ⣽⬊ෆ௦ㅰ㓟≀ࡀ࢚ࣆࢪ࢙ࢿ ࢸࢵࢡไᚚࢆࡶྖࡗ࡚࠸ࡿࡇࡶᚎࠎ᫂ࡽ ࡞ࡗ࡚ࡁ࡚࠾ࡾᢠ㓟స⏝ࡢࡳ࡛࡞ࡃᚋࡑࡢస⏝ 㝿࡞࣓࢝ࢽࢬ࣒ゎᯒࡘࡁ᳨ウࢆࡍࡍࡵ࡚࠸ࡃணᐃ ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 422 - බເᨭㄢ㢟㸸2010 ᖺᗘ ࣃ࣮ࢯࢼࣝࢤࣀ࣒ゎᯒᇶ࡙ࡃ $/6 ࡢᝈ㛵㐃㑇ఏᏊ᥈⣴ែゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ ⏣୰ ❶ᬒ࣭ྡྂᒇᏛᏛ㝔་Ꮫ⣔◊✲⛉ ⌧ᡤᒓ㸹ᶓᕷ❧ᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ➽ⴎ⦰ᛶഃ⣴◳㸦ALS㸧ࡣࠊ㐠ືࢽ࣮ࣗࣟࣥ ࡀ≉␗ⓗ㞀ᐖࡉࢀኚᛶ⬺ⴠࡍࡿ⚄⤒ኚᛶᝈ࡛࠶ ࡾࠊ㐍⾜ᛶࡢ➽ⴎ⦰➽ຊపୗࢆࡁࡓࡋࠊⓎࡽ ᖹᆒ 3-4 ᖺ࡛Ṛஸࡲࡓࡣேᕤ྾ჾ╔ࡀᚲせ࡞ ࡿᝈ࡛࠶ࡿࠋALS ࡣࣝࢶࣁ࣐࣮ࡸࣃ࣮࢟ࣥ ࢯࣥẚ࡚ᝈ⪅ᩘࡣᑡ࡞࠸ࡶࡢࡢࠊࡑࡢ㔜⠜ࡉ ᝈ⪅࠾ࡼࡧᐙ᪘ㄢࡏࡽࢀࡿ㐣㓞࡞㐠ࡽࠊ㞴 ୰ࡢ㞴ゝࢃࢀ࡚࠾ࡾࠊࡑࡢඞ᭹ࡣႚ⥭ࡢㄢ㢟 ࡞ࡗ࡚࠸ࡿࠋᡃࠎࡣࠊALS ࡢែࢆࢤࣀ࣒ゎᯒ ࡽ᫂ࡽࡍࡿࡇࢆ┠ᣦࡋࠊᝈ⪅ DNA 㞟✚ࡢࡓࡵ ࡢࢩࢫࢸ࣒ᵓ⠏ࢆ᥎㐍ࡋ࡚ࡁࡓࠋ2006 ᖺࡼࡾྡྂᒇ Ꮫ⚄⤒ෆ⛉ࢆ୰ᚰ 30 タ௨ୖࡀཧຍࡍࡿᅜ つ ᶍ ࡢ Japanese Consortium for Amyotrophic Lateral Sclerosis Research (JaCALS)ࢆ❧ࡕୖࡆࠊ DNAࠊᰴ⣽⬊ࠊ⤒ⓗ⮫ᗋሗࡢ㞟ࢆ⾜ࡗ࡚ࡁ ࡚࠸ࡿࠋࡍ࡛ 1000 ࢆ㉸࠼ࡿ JaCALS ᳨యࣜࢯ ࣮ࢫ๓ྥࡁ⮫ᗋሗࢆ㞟✚ࡋ࡚࠾ࡾࠊࡇࢀࡽࢆ⥙ ⨶ⓗゎᯒࡋ ALS ࡢែゎ᫂ࠊ⒪ἲ㛤Ⓨࢆ┠ᣦࡍࠋ 㸺ᨭࡢෆᐜ㸼 ALS ࡢࢇࡀᏙⓎᛶ࡛࠶ࡾࠊᐙ᪘ᛶࡢࡶࡢࡀ ༨ࡵࡿྜࡣ 5-10%Ỵࡋ࡚㧗ࡃࡣ࡞࠸ࠋࡋࡋࠊ ㏆ᖺྠᐃࡉࢀࡓᐙ᪘ᛶ ALS ࡢᅉศᏊ TAR DNA binding protein 43 kDa(TDP-43)ࡣࠊᏙⓎᛶ ALS ࡢ 㐠ືࢽ࣮ࣗࣟࣥ࠾࠸࡚ࠊࡑࡢ⣽⬊ෆᒁᅾࡀ᰾ෆ ࡽ⣽⬊㉁ኚࡋࣘࣅ࢟ࢳࣥ㝧ᛶᑒධయࡢせᵓ ᡂᡂศ࡞ࡗ࡚࠸ࡿࡇࡀ᫂ࡽࡉࢀࡿ࡞ࠊᏙ Ⓨᛶ ALS ࡗ࡚ࡶᴟࡵ࡚㔜せ࡞ែ㛵㐃ศᏊ࡛࠶ ࡿࡇࡀ♧ࡉࢀ࡚࠸ࡿࠋ᪂ࡓᐙ᪘ᛶ ALS ࡢᅉ㑇 ఏᏊࢆྠᐃࡍࡿࡇࡣࠊᏙⓎᛶ ALS ゎ᫂ࡢࡓࡵࡶ 㠀ᖖ㔜せ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋࡑࡇ࡛ࠊᡃࠎࡀ㑇 ఏᏊ᳨యࡸ⮫ᗋሗࢆ㞟✚ࡋ࡚ࡁࡓᐙ᪘ᛶ ALS ࡢ 6 ᐙ⣔ࠊᝈ⪅࠾ࡼࡧ㠀Ⓨ⪅ࢆ࠶ࢃࡏࡓ 11 ྡࡼࡾ DNA ࡢᥦ౪ࢆཷࡅࠊᮏࢤࣀ࣒ᨭࡼࡿࢤࣀ࣒ゎᯒࡢྠ ពࢆᚓ࡚ࠊᖹᡂ 22 ᖺᗘࡢࢤࣀ࣒ᨭࡼࡾḟୡ௦ࢩ ࣮ࢡ࢚ࣥࢧ࣮ࢆ⏝࠸ࡓ࢚ࢡࢯ࣮࣒ゎᯒࢆ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ゎᯒࢆ⾜ࡗࡓ 11 ྡ࠾࠸᳨࡚ฟࡉࢀࡓ SNV ࡢ ᩘࡣᖹᆒࡋ࡚ྛࠎ⣙ 450,000 ⛬ᗘ࡛࠶ࡾࠊࡇࡢ࠺ࡕ ᪤▱ࡢከᆺࢆ㝖እࡋࡓ rare ࡞ࡶࡢࡀ⣙ 30,000 ⛬ᗘࠊ ࡑࡋ࡚ࡑࡢ࠺ࡕࠊ㠀ྠ⩏⨨ࡸࢫࣉࣛࢫ㒊ࡢኚ ␗ࡣ⣙ 400 ⛬ᗘ᫂ࡽ࡞ࡗࡓࠋ୍᪉ࠊindel ࡘ࠸ ࡚ࡣࠊ⣙ 35,000 ⛬ᗘࡀྠᐃࡉࢀࠊࡑࡢ࠺ࡕ᪤▱ࡢከ ᆺࢆ㝖እࡋࡓ rare ࡞ࡶࡢࡀ⣙ 2,500ࠊࢥ࣮ࢹࣥࢢ 㡿ᇦࡢ࠶ࡿࡶࡢࡀ⣙ 35 ๓ᚋ࡛࠶ࡗࡓࠋࡑࡋ࡚ࠊྛࠎ ࡢᐙ⣔ࡘ࠸࡚ࠊ㑇ఏᵝᘧࠊⓎ⪅࣭㠀Ⓨ⪅ࡼ ࡿ⤠ࡾ㎸ࡳࢆ⾜ࡗࡓࠋࡇࡢ⤖ᯝࠊᐙ⣔ࡼࡗ࡚㐪࠸ ࡣ࠶ࡿࡀࠊࢧࣥࣉࣝᩘࡀẚ㍑ⓗ☜ಖ࡛ࡁ࡚࠸ࡿᐙ⣔ ࡘ࠸࡚ࡣᅉ㑇ఏᏊೃ⿵ࢆ SNV ࡛ᩘ༑࢝ᡤࠊindel ࡛ᩘ࢝ᡤࡲ࡛⤠ࡾ㎸ࢇࡔࠋ ᨭࡼࡾᚓࡽࢀࡓ⤖ᯝࢆⓎᒎࡉࡏࡿࡃࠊᨭ ⤊ᚋࡶᐙ⣔ෆࡢࢧࣥࣉࣝ㞟✚ࢆ㐍ࡵࡓࡀࠊࡍ࡛ Ṛஸࡋ࡚࠸ࡓࡾࠊ༠ຊࡀᚓࡽࢀ࡞࠸࡞ࡢၥ㢟ࡢࡓ ࡵࠊࡉࡽ࡞ࡿ⤠ࡾ㎸ࡳࡀ࡛ࡁࡿࡇࢁࡣ⮳ࡗ࡚࠸ ࡞࠸ࠋࡢᐙ᪘ᛶ⚄⤒ᝈ␗࡞ࡾࠊALS ࡣⓎ ࡽṚஸࡲ࡛ࡢᮇ㛫ࡀ▷ࡃᐙ⣔ෆⓎ⪅ࡀࡍ࡛Ṛஸ ࡋ࡚࠸ࡿࡇࡀከࡃࠊࡲࡓᴟࡵ࡚㔜⠜࡞ᝈ࡛࠶ࡿ ࡓࡵࠊ༠ຊࢆ౫㢗ࡋ࡚ࡶࠊᐙ᪘ᛶ࡛࠶ࡿࡇࡀྠ⬊ ఏࢃࡿࡢࢆᜍࢀ࡚ᛮ࠺ࡼ࠺ࢧࣥࣉࣝ㞟ࡀ㐍ࡲ ࡞࠸࠸ࡗࡓ㑇ఏᛶ㞴ᛶᝈ≉᭷ࡢၥ㢟ࡀ࠶ࡿࠋ ᐙ᪘ᛶ ALS ࡛ࡣࠊࡑࡢᅉ㑇ఏᏊࡶ heterogeneous ࡛࠶ࡿࡇࡽࡶࠊᚋࡶ᪂ࡓ࡞ᐙ⣔ࢆⓎ᥀ࡋ࡚ゎ ᯒࡍࡿࡇ࡛ࠊ」ᩘᐙ⣔ඹ㏻ࡍࡿኚ␗ࢆぢฟࡍࡇ ࡀ࡛ࡁࢀࡤ⪃࠼࡚࠸ࡿࠋᨭࢆཷࡅ࡚ࡽࡍ࡛ 4 ᖺࡀ⤒㐣ࡋ࡚࠸ࡿࡀࠊࡑࡢࢹ࣮ࢱࡣ༑ศά⏝ ࡉࢀ࡚࠾ࡾࠊᚋࡶ᪂ࡓ࡞ ALS ཎᅉࡲࡓࡣ㛵㐃㑇ఏ Ꮚࡢྠᐃࡘ࡞ࡀࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᡃࠎࡀᨭࢆཷࡅࡓᖹᡂ 22 ᖺᗘᙜࡣࠊ࢚ࢡࢯ࣮ ࣒ࢩ࣮ࢡ࢚ࣥࢫせࡍࡿࢥࢫࢺࡣ⌧ᅾࡼࡾࡶ㐭 㧗㢠࡛࠶ࡾࠊಶࠎࡢ◊✲ᐊࡢ㝈ࡽࢀࡓண⟬࠾࠸࡚ ࢚ࢡࢯ࣮࣒ࢩ࣮ࢡ࢚ࣥࢫࢆ⾜࠺ࡇࡣࡁ࡞ᅔ㞴 ࢆకࡗࡓࠋᝈࡢ≉␗ᛶࡶᙳ㡪ࡋࠊ᭱⤊ⓗᙜึ┠ ᣦࡋ࡚࠸ࡓᐙ᪘ᛶ ALS ᪂つཎᅉ㑇ఏᏊࡢྠᐃࡲ࡛ ࡣ⮳ࡗ࡚࠸࡞࠸ࡀࠊᚋ᪂ࡓ࡞ᐙ⣔ࡢゎᯒࡼࡾᚓ ࡽࢀࡓࢹ࣮ࢱẚ㍑᳨ウࡍࡿࡇ࡛ࠊ᪂ࡓ࡞Ⓨぢ ࡘ࡞ࡀࡿྍ⬟ᛶࡀ࠶ࡿࠋࡉࡽࠊࡇࢀࡽࡢ⤒㦂ࢆࡶ ࠊ⌧ᅾࠊᏙⓎᛶ ALS 㛵ࡋ࡚ࡶ࢚ࢡࢯ࣮࣒ࢩ࣮ ࢡ࢚ࣥࢫゎᯒࢆ᥎㐍ࡋ࡚࠸ࡿࠋ⌧Ⅼ࠾࠸࡚ 600 ௨ୖࡢ࢚ࢡࢯ࣮࣒ࢩ࣮ࢡ࢚ࣥࢫࡀ⤊ࡋ࡚࠾ࡾࠊ 㡰ḟࢹ࣮ࢱゎᯒࢆ㐍ࡵ࡚࠸ࡿࡇࢁ࡛࠶ࡿࠋ࢚ࢡࢯ ࣮࣒ࢩ࣮ࢡ࢚ࣥࢫࢹ࣮ࢱ࣮ࢆࡶᵓ⠏ࡉࢀࡓ᪥ᮏ ேṇᖖࢥࣥࢺ࣮ࣟࣝࡢࢹ࣮ࢱ࣮࣋ࢫ࡛࠶ࡿ HGVDࠊ ࡉࡽࡣࢤࣀ࣒ࢩ࣮ࢡ࢚ࣥࢫࢹ࣮ࢱࢆ࣮࣋ࢫࡋ ࡓᮾ࣓ࢹ࣭࣓࢝ࣝ࢞ࣂࣥࢡᶵᵓࡀබ㛤ࡋ࡚࠸ࡿ ᪥ᮏேṇᖖࢥࣥࢺ࣮ࣟࣝࢹ࣮ࢱࡢẚ㍑᳨ウࢆ⾜࠸ ᏙⓎᛶ ALS ࡢⓎࡢཎᅉ࡞ࡿኚ␗ࢆゎᯒࡋ࡚࠸ࡿࠋ ࡲࡓࠊ๓ྥࡁ⮫ᗋሗࢆά⏝ࡋࡓ ALS ែ㛵㐃㑇ఏ Ꮚࡢ᥈⣴࡞ࠊALS ែゎ᫂ྥࡅࡓࢤࣀ࣒◊✲ࡀ ⌧ᅾࡶ㐍⾜୰࡛࠶ࡿࠋ ᨭࢆ㏻ࡌ࡚ࠊ࢚ࢡࢯ࣮࣒ゎᯒࡢጾຊࢆᐇឤࡍࡿ ࡶࢧࣥࣉࣝ㞟ࡢ㔜せᛶࢆㄆ㆑ࡍࡿࡇࡀ ࡛ࡁࠊࢤࣀ࣒◊✲᥎㐍ྥࡅ࡚ࡢᇶ┙ࢆᵓ⠏ࡍࡿࡇ ࡀ࡛ࡁࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Nakamura, R. et al., Neurobiol Aging. 2015 Dec 7. [Epub ahead of print] - 423 - බເᨭㄢ㢟㸸2013 ᖺᗘ ᳜≀ඹ⏕ࡍࡿ࣓ࢱࣀ࣮ࣝ㈨ᛶ⣽⳦ࡢከᵝᛶඹ⏕ᶵᵓࡢゎ᫂ ⱝᡭ◊✲㸦㸿㸧 ㇂ ᫂⏕࣭ᒸᒣᏛ㈨※᳜≀⛉Ꮫ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 Methylobacterium ᒓ⣽⳦ࡣ᳜≀ࡀᨺฟࡍࡿ࣓ࢱ ࣀ࣮ࣝࢆ⏝ࡋ࡚᳜≀⾲㠃⣽⳦ࡢ 10-20%ࢆ༨ࡵࡿ ඃ༨ࡋࠊ᳜≀ࡢ⏕⫱ࢆಁ㐍ࡍࡿࠋಁ㐍㛵 ࡍࡿࡉࢀࡿ᳜≀࣍ࣝࣔࣥࡢྜᡂ⬟ࠊ↓ᶵ≀㸦ࣜࣥࠊ 㕲➼㸧ࡢ౪⤥⬟ࡀ࠶ࡿࡇࡀάᛶ࡛ࣞ࣋ࣝศࡗ࡚ ࠸ࡿࡀࠊಁ㐍ࡢヲ⣽࡞࣓࢝ࢽࢬ࣒ࡣࡼࡃศࡗ࡚࠸ ࡞࠸ࠋࡲࡓᮏᒓ⣽⳦ࡣ࣓ࢱࣀ࣮ࣝࢆ㓟ࡍࡿ㓝⣲ࠊ ࣓ࢱࣀ࣮ࣝ⬺Ỉ⣲㓝⣲㸦MDH㸧ࢆࡘࡶࡕࠊࡑࢀࡒ ࢀ࢝ࣝࢩ࣒࢘ࣛࣥࢱࣀࢻඖ⣲౫Ꮡࡍࡿ㓝⣲࡛ ࠶ࡿࡇࡀศࡗ࡚࠸ࡿࠋࡇࡢ୧⪅ࡢ㔠ᒓ౫Ꮡࡋ ࡓⓎ⌧ࡸࡘᏑᅾࡍࡿព⩏ࡣࡃศࡗ࡚࠸࡞࠸ࠋ 㸺ᨭࡢෆᐜ㸼 ᮏㄢ㢟࡛ࡣ⏦ㄳ⪅ࡀศ㞳ࡋࡓࠊ࢚ࢰࢫࢼࢦࢣࡽࡢ ศ㞳⳦ M. aquaticum 22A ᰴࡢࢻࣛࣇࢺࢤࣀ࣒ሗ ࢆࣇࢽࢵࢩࣥࢢࡋ࡚ࠊ᳜≀ࡢ┦స⏝㔜せ࡞ 㑇ఏᏊࡢࢤࣀ࣒ୖࡢ⨨㛵ಀࢆࡣࡗࡁࡾࡉࡏࠊᚋ ࡢᶵ⬟ゎᯒ⏝ࡍࡿࠋࡲࡓࠊ22A ᰴࢆࣔࢹࣝࡋ ࡚࢝ࣝࢩ࣒࢘ࣛࣥࢱࣀࢻᏑᅾୗ࡛࣓ࢱࣀ࣮ࣝ ⏕⫱ࡋࡓ㝿ࡢ⥙⨶ⓗࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢆ⾜࠸ࠊ 㔠ᒓࡢᏑᅾࡼࡾኚࡍࡿ࣓ࢱࣀ࣮ࣝ௦ㅰ⤒㊰㑇ఏ Ꮚࡸࡑࡢไᚚ㛵ࢃࡿᅉᏊࡢྠᐃ⤖ࡧࡘࡅࡿࠋࣔ ࢹ᳜ࣝ≀ࡋ࡚ࢩࣟࢾࢼࢬࢼࢆ⏝࠸ࠊ22A ᰴඹ ⏕ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࡶィ⏬ࡋࡓࡀ RNA 㔞 ࡀ㊊ࡋࡓࡓࡵᐇ㦂ࢆ⾜ࡗ࡚࠸࡞࠸ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ձ22A ᰴࢤࣀ࣒ࡢࣇࢽࢵࢩࣥࢢ ࡍ࡛ᚓࡽࢀ࡚࠸ࡓࢻࣛࣇࢺࢤࣀ࣒ࡣ contig ᩘࡀ 500 ᑍ᩿ࡉࢀ࡚࠾ࡾࠊ㑇ఏᏊࡢ⨨㛵ಀࢆᥗࡳ ࡡ࡚࠸ࡓࠋ454 Illumina ࡢ࣮ࣜࢻࢆຍ࠼ࠊ47x ࢝ ࣂ࣮ࣞࢪࡋࠊࢠࣕࢵࣉࢆ PCR ࡛ᇙࡵ࡚࠸ࡓࡔࡁࣇ ࢽࢵࢩࣥࢢ㡬࠸ࡓࠋ ࢺ࣮ࢱࣝࢤࣀ࣒ࡣ 7.56 Mbࠊࣉࣛࢫ࣑ࢻࡣ㸳ࡘ࡛ ࠶ࡿࡇࡀศࡗࡓࠋPGAP, MiGAP ࡼࡿࣀࢸ ࣮ࢩࣙࣥࢆ⾜࠸ 6944 ࡢ CDS ࢆぢ࠸ࡔࡋࡓࠋࣜ࣎ࢰ ࣮࣒ RNA ࢜࣌ࣟࣥࡣ 11ࠊtRNA 㑇ఏᏊࡣ 95 ಶᏑᅾ ࡋࡓࠋMDH ࢆࡣࡌࡵ࣓ࢱࣀ࣮ࣝ௦ㅰ㛵ࢃࡿ㓝⣲㑇 ఏᏊ⩌ࠊPQQ ྜᡂ㓝⣲㑇ఏᏊࠊࢧࢺ࢝ࢽࣥ 㸦t-zeatin㸧ྜᡂ㑇ఏᏊࠊ࢚ࢳࣞࣥྜᡂࡢ㜼ᐖ㛵ࢃ ࡿ ACC deaminase 㑇ఏᏊ, ࢩࢹࣟࣇ࢛ྜᡂ㑇ఏ Ꮚࠊࡲࡓ㐠ືᛶ㛵ࢃࡿࣇࣛࢪ࢙ࣜࣥ㑇ఏᏊࠊType VI ศἪ⣔㑇ఏᏊ➼ࡀぢ࠸ࡔࡉࢀࡓࠋ࡞࠾❅⣲ᅛᐃࠊ Type III, IV ศἪ⣔ࡣぢ࠸ࡔࡉࢀ࡞ࡗࡓࠋ ࢱࣀ࣮ࣝࠊཬࡧࢥࣁࢡ㓟ࢆ⏝࠸࡚⏕⫱ࡋࡓ⳦య RNA ࢆ⏝࠸࡚࡚ 3 㐃࡛ RNA-Seq ゎᯒࢆ⾜ࡗ࡚࠸ࡓࡔ ࠸ࡓࠋ࡞࠾ RNA ࡣ riboZero ࡼࡾࣜ࣎ࢰ࣮࣒ RNA ࢆ㝖࠸ࡓࠋ࣮ࣜࢻࢹ࣮ࢱࢆ㑇ఏ◊ࡢࢧ࣮ࣂ࣮࠶ࡿ Maser ࡛ゎᯒࡋࡓࠋ࣮ࣜࢻࡣձࡼࡗ࡚ᚓࡽࢀࡓࢤ ࣀ࣒ BWA ࡛࣐ࢵࣆࣥࢢࡋࡓࠋrRNA ࣐ࢵࣉࡉࢀ ࡓ࣮ࣜࢻࢆ㝖ࡁ CDS ࡈࡢ FPKM ࢆィ⟬ࡋࡓࠋ ࡲࡎὀ┠ࡉࢀࡓࡢࡣ࢝ࣝࢩ࣒࢘(Ca)౫Ꮡࡢ MDH ࡛࠶ࡿ MxaF ࣛࣥࢱࣀࢻ(La)౫Ꮡࡢ XoxF ࡛࠶ ࡗࡓࠋࡇࢀࡽࡣࡑࢀࡒࢀࡢ㔠ᒓࡢᏑᅾ౫Ꮡ ࡋ࡚Ⓨ⌧ࡋ࡚࠾ࡾࠊࡲࡓ୧㔠ᒓࡀᏑᅾࡍࡿ XoxF ࡢⓎ⌧ࡀඃඛࡋ࡚࠸ࡿࡇࡽࠊᮏᒓ⣽⳦ࡣࣛࣥࢱ ࣀࢻࢆዲࢇ࡛⏝ࡋ࡚࠸ࡿࡇࡀ࠺ࡀ࠼ࡓࠋ MDH ࡢ⏘≀࡛࠶ࡿ࣒࣍ࣝࣝࢹࣄࢻ௨㝆ࡢ௦ㅰ 㛵ࢃࡿ㑇ఏᏊࡢⓎ⌧㛵ࡋ࡚ࡣࢇኚࢃࡽࡎࠊ 㔠ᒓࡣ࣓ࢱࣀ࣮ࣝ㓟ࡢࡳ㛵ࢃࡗ࡚࠸ࡿࡇ ࡀศࡗࡓࠋMxaF ࡣ Ca ࢆศᏊෆ⤖ྜࡍࡿࡓࡵࡢ ከࡃࡢࢱࣥࣃࢡ㉁ࢆᚲせࡋ࡚࠾ࡾࠊࡑࢀࡽࡀ Ca Ꮡᅾୗ࡛ㄏᑟࡉࢀ࡚࠸ࡿࡇࡶぢ࡚ྲྀࢀࡓࠋࡲࡓ La ࡛≉␗ⓗⓎ⌧㔞ࡢ㧗࠸㑇ఏᏊࡢከࡃࡢᶵ⬟ࡣ᫂ ࡞ࡶࡢࡀከࡗࡓࠋ ◊✲ࡢࣔࢹࣝࡋ࡚⏝࠸࡚࠸ࡿ 22A ᰴࡢࢤࣀ࣒ࡀ ᫂ࡽ࡞ࡾࠊRNA-Seq ゎᯒࢹ࣮ࢱࡣ࣓ࢱࣀ࣮ࣝ ࡑࢀ௨እࡢⅣ⣲※ࡢ⏕⫱ࡢࡁ࡞㑇ఏᏊⓎ⌧ኚ ࠊCa,La ࡼࡿ≉␗ⓗ࡞ኚࡀぢࡽࢀࠊᚋࡢ ◊✲㠀ᖖ᭷┈࡞ሗᇶ┙࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ⳦ࢤࣀ࣒ࡢ GC ྵ㔞ࡣᖹᆒ 69.1%࡛࠶ࡿࡀࠊLa ࡛㧗Ⓨ⌧ࡍࡿ CDS ࡣ GC ྵ㔞ࡢప࠸(<60%)CDS ࡀከ࠸ࡇࡀศࡗࡓࠋࡇࢀࡽࡣ GC ྵ㔞ࡀࡁࡃ ␗࡞ࡿࡇࡽእ᮶㑇ఏᏊ࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋLa ࡢᏑᅾࡀ RNA-Seq ࡢゎᯒ⮬యᙳ㡪ࡋࡓࡢ࡛࡞ࡅ ࢀࡤࠊᮏᒓ⣽⳦ࡢࢤࣀ࣒ࢥ࣮ࢻࡉࢀࡿእ᮶㑇ఏᏊ ࡣ࣍ࢫࢺ࡞ࡿ᳜≀ෆ࡛㔜せ࡛࠶ࡿ࠸࠺ሗ࿌ࡶ࠶ ࡿࡇࡽࠊࡑࢀࡽࡀ La Ꮡᅾୗ࡛㧗Ⓨ⌧ࡍࡿࡇࡣ ᳜≀ࡢඹ⏕ࡶఱࡽࡢពࡀ࠶ࡿ⪃࠼ࡽࢀࡓࠋ ᚋ⌧ᛶ transcript ᵓ㐀ࢆ᫂ࡽࡋ࡚ࢱࣥࣃ ࢡ㉁ࢆࢥ࣮ࢻࡋ࡚࠸ࡿ࠺࡞᳨ウࡋ࡚࠸ࡃண ᐃ࡛࠶ࡿࠋࡇࡢࡼ࠺ࢤࣀ࣒ᨭࠊRNA-Seq ᨭ ࡼࡗ࡚᪂つ࡞⌧㇟ࡢ genome, transcriptome ࢹ࣮ࢱ ᇶ┙ࡀ࡛ࡁ࠶ࡀࡾࠊᕼᅵ㢮ᛂ⟅㑇ఏᏊ᳜≀ࡢඹ ⏕ࡢ㛵ࢃࡾࡢゎᯒ᪂ࡓ࡞㊊ࡀࡾࡀᚓࡽࢀ࡚࠸ ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ղ࢝ࣝࢩ࣒࢘ࠊࣛࣥࢱࣀࢻᏑᅾୗ࣓ࢱࣀ࣮ࣝ⏕⫱ ࡢ 22A ᰴࡢ RNA-Seq ゎᯒ ࢝ࣝࢩ࣒࢘ཬࡧࣛࣥࢱࣀࢻࡋ࡚ࠊࣛࣥࢱࣥࠊ ࣑࣒࣍ࣝ࢘ࠊࣝࢸࢳ࣒࢘ࡢᏑᅾୗ࡛Ⅳ⣲※ࡋ࡚࣓ Tani A., et al., Genome Announc 3(2) e00266-15 (2015) - 424 - බເᨭㄢ㢟㸸2010 ᖺᗘ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ࡢグ᠈ᙧᡂᅇ㊰ࡢᵓ㐀࠾ࡼࡧᶵ⬟Ⓨ⌧ࡢศᏊᇶ┙ ᪂Ꮫ⾡㡿ᇦ◊✲ ከ⩚⏣ ဴஓ࣭ᮾிᏛศᏊ⣽⬊⏕≀Ꮫ◊✲ᡤ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚グ᠈ᙧᡂ㐣⛬࡛ࡣ cAMP ⤒㊰ࡀ せ࡞ᙺࢆᣢࡘ⪃࠼ࡽࢀ࡚࠾ࡾࠊࡇࢀࡲ࡛ Adenylyl Cyclase (rutabaga 㑇ఏᏊ)ࡸ PKA ࡞ ࡘ࠸࡚ヲ⣽࡞ゎᯒࡀ⾜ࢃࢀ࡚ࡁࡓࠋᮏ◊✲࡛ࡣࠊ cAMP ⤒㊰ࢆグ᠈ᙧᡂࡢ୰ᚰ㐣⛬ぢ❧࡚ࠊ㛗ᮇ グ᠈ᙧᡂ cAMP ⤒㊰ࢆಟ㣭ࡍࡿྍ⬟ᛶࡢ࠶ࡿ ᅉᏊࢆㄪࡓࠋPhosphodiesterase㸦PDE㸧ࢆࢥ࣮ ࢻࡋ࡚࠸ࡿ dunce 㑇ఏᏊࡣ cAMP 㔞ࡢㄪ⠇ࢆࡋ ࡚グ᠈ᙧᡂ㛵ࡋ࡚࠸ࡿ⪃࠼ࡽࢀ࡚࠸ࡿࡀࠊ 㛗ᮇࡔࡅ࡛ࡣ࡞ࡃ▷ᮇグ᠈ᙧᡂࡶാࡃࠋࢩࣙ ࢘ࢪࣙ࢘ࣂ࢚ࡣᑡ࡞ࡃࡶ 6 ࡘࡢ PDE ࢱࣥࣃ ࢡࡀᏑᅾࡍࡿࡀࠊDunce ௨እࡘ࠸࡚ࡣ⚄⤒ᶵ⬟ 㛵ࡍࡿ࠺ࡣ࡛᫂࠶ࡿࠋࡑࡇ࡛ࢩࣙ࢘ 㸺ᨭࡢෆᐜ㸼 ࢪࣙ࢘ࣂ࢚ PDE 㑇ఏᏊࡘ࠸࡚࢟ࣀࢥయ࡛ࡢⓎ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ᑐࡋ࡚ໝ࠸㟁Ẽ่⃭ࢆ⧞ࡾ ⌧㔞ࢆࢩ࣮ࢡ࢚ࣥࢩࣥࢢࡢ⤖ᯝࢆ⏝࠸࡚ẚ㍑ࡋࡓ ㏉ࡋᥦ♧ࡍࡿࡇࡼࡾࠊ㛗ᮇႥぬᚷ㑊グ᠈ ࡇࢁ pde1c ࡀ࢟ࣀࢥయ࡛Ⓨ⌧ࡍࡿࡇࡀࢃࡗ 㸦long-term-olfactory-aversive-memory㸧ࢆᙧᡂ ࡉࡏࡿࠋ୍ᐃ㛫⨨࠸ࡓᚋゎ๗ࡋ࢟ࣀࢥయࢆ ࡓࠋin situ hybridization ࢆ⾜ࡗࡓࡇࢁ pde1c ฟࡍࡿࠋTotal RNA ࢆ࢟ࣀࢥయࡽᢳฟࡋࠊ㏫㌿ ࡀ࢟ࣀࢥయ࡛ᙉࡃⓎ⌧ࡍࡿࡇࢆ☜ㄆ࡛ࡁࡓࠋࡉ ࣭PCR ቑᖜࡼࡾᮏ㙐 cDNA ࢆసᡂࡍࡿࠋ ࡽ pde1c 㑇ఏᏊ㡿ᇦࡢࢺࣛࣥࢫ࣏ࢰࣥᤄධ⣔⤫ ᡂࡋࡓ cDNA ࢆࢩ࣮ࢡ࢚ࣥࢩࣥࢢࡍࡿࡇࡼࡾࠊ ࢆ⏝࠸࡚グ᠈ࢵࢭࢆ⾜ࡗࡓ⤖ᯝࠊ▷ᮇグ᠈ࡣ ࢟ࣀࢥయ࡛Ⓨ⌧ࡍࡿ㑇ఏᏊࢆ⥙⨶ⓗㄪࡿࠋࡉ ṇᖖ࡞୍᪉࡛ࠊspaced training ᚋ㸰㸲㛫࠾ࡅ ࡽࠊᏛ⩦๓ᚋ࡛ẚ㍑ࡍࡿࡇࡼࡾࠊ࢟ࣀࢥయ ࡿ㛗ᮇグ᠈ᙧᡂࡀᙉࡃᢚไࡉࢀ࡚࠸ࡓࠋࡇࡢࡇ ࠾࠸࡚グ᠈ᙧᡂ౫Ꮡⓗୖ᪼ࡍࡿ㑇ఏᏊࢆ⥙⨶ ࡣࠊ㛗ᮇグ᠈ᙧᡂ≉␗ⓗാࡃࡼ࠺࡞ cAMP ⓗㄪࡿࠋ㛗ᮇႥぬグ᠈ࡢ᮲௳ࡅࡣࠊ㛫㝸 ⤒㊰ไᚚᶵᵓࡢᏑᅾࢆ♧၀ࡋ࡚࠸ࡿࠋpde1c 㑇ఏ ࢆ✵ࡅ่࡚⃭ᥦ♧ࢆ⧞ࡾ㏉ࡍ spaced training Ꮚグ᠈⾲⌧⣔ࡢᑐᛂ㛵ಀࡘ࠸࡚ࡉࡽゎᯒ 㛫㝸ࢆ✵ࡅࡎ⧞ࡾ㏉ࡍ massed training ࡀ࠶ࡾࠊ ࢆࡍࡍࡵࡿࡇ࡛ࠊグ᠈ࡀᅛᐃࡉࢀࡿᶵᵓࡘ ๓⪅ࡼࡾ CREB ౫Ꮡᛶࡢ㛗ᮇグ᠈ࡀᙧᡂࡉࢀࡿࠋ ࠸࡚᪂ࡓ࡞▱ぢࡀᚓࡽࢀࡿᮇᚅࡉࢀࡿࠋ ᮏ◊✲࡛ࡣ spaced sample massed sample ࢆẚ ᮏࣉࣟࢪ࢙ࢡࢺࡼࡗ࡚⥙⨶ⓗྠᐃࡉࢀࡓ㑇ఏ ㍑ࡍࡿࡇࡼࡾࠊ㛗ᮇグ᠈≉␗ⓗ࡞㑇ఏᏊኚ Ꮚ⩌ࡣ㐣ཤࡢሗ࿌ࡣᚲࡎࡋࡶ୍⮴ࡋ࡞࠸ࠋ㛗ᮇグ ືࢆᢳฟࡋࡼ࠺⪃࠼ࡓࠋ ᠈ᙧᡂࡢᡭἲࡀᚲࡎࡋࡶྠ୍࡛ࡣ࡞࠸ࡇ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚㛗ᮇႥぬグ᠈ᙧᡂࡣ᪂ࡓ࡞㑇 ఏᏊⓎ⌧ࡀᚲせ࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋᮏ◊ ✲࡛ࡣձ㛗ᮇグ᠈ᙧᡂ㌿άᛶࡉࢀࡿᅉᏊ ࡢྠᐃࠊղグ᠈ᙧᡂࡢ୰ᚰⓗ࡞ศᏊ⤒㊰࡛࠶ࡿ cAMP ⤒㊰ࢆ㛗ᮇグ᠈ᙧᡂ≉␗ⓗಟ㣭ࡍࡿศᏊ ᶵᵓࡢゎ᫂ࠊࡢ㸰Ⅼ↔Ⅼࢆࡋࡰࡾࠊ㛗ᮇグ᠈ᙧ ᡂᶵᵓࢆゎ᫂ࡍࡿࡇࢆ┠ⓗࡋ࡚࠸ࡿࠋලయⓗ ࡣࢩࣙ࢘ࢪࣙ࢘ࣂ࢚Ⴅぬグ᠈࠾࠸࡚୰ᚰⓗ࡞ ᙺࢆᯝࡓࡍ࢟ࣀࢥయ⚄⤒⣽⬊࡛グ᠈ࡼࡾⓎ⌧ ኚືࡍࡿ㑇ఏᏊࡢྠᐃࢆ⾜࠺ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡇࢀࡲ࡛ࢩ࣮ࢡ࢚ࣥࢩࣥࢢࡼࡾࠊᏛ⩦ᚋ㸯㸰 㛫ࡢ spaced sample㸻㸱࣌ࠊᏛ⩦ᚋ㸴㛫ࡢ spaced sample㸻㸰࣌ࠊᏛ⩦ᚋ㸴㛫ࡢ massed sample㸻㸰࣌ࠊ࠾ࡅࡿⓎ⌧ゎᯒࢆ⾜ࡗࡓ㸦Ꮫ ⩦ࡋࡓࢧࣥࣉࣝࠊఱࡶ่⃭ࢆຍ࠼࡚࠸࡞࠸ࢥࣥ ࢺ࣮ࣟࣝࡢࢧࣥࣉࣝࢆ୍ᑐࡋ࡚ࠊゎᯒࡢࠕ࣌ ࠖࡪ㸧ࠋ12 㛫ᚋࡢࢧࣥࣉࣝࡘ࠸࡚ࡣࠊ ༑ศ࡞ࢱࢢᩘࡀᚓࡽࢀ࡞ࡗࡓࡓࡵࠊ㸴㛫ᚋࡢ ࢧࣥࣉࣝࢆゎᯒࡋࡓࠋࡲࡎ㸴㛫ᚋࡢ spaced sample ࠾࠸࡚ձⓎ⌧ୖ᪼ࡋ࡚࠸ࡿࠊղⓎ⌧ࣞ࣋ ࣝࡀ㧗࠸ࠊճグ᠈ᙧᡂ㛵ࢃࡿࡇࡀ▱ࡽࢀ࡚࠸ ࡿ⺮ⓑ㉁ࡢᶵ⬟ࢻ࣓ࣥࢆࡶࡘࠊ࠸࠺᮲௳ࢆタ ᐃࡋ㑇ఏᏊࢆᢳฟࡋࡓࠋࡉࡽࡑࢀࡽࡢ㑇ఏᏊ ࡘ࠸࡚ࠊմmassed sample ࡛ῶᑡࡋ࡚࠸ࡿ࠶ࡿ࠸ ࡣኚࡋ࡞࠸ࠊࡶࡢࢆᢳฟࡋ㸱㸮㑇ఏᏊࢆ≉ᐃࡋ ࡓ ࠋ ࡇ ࢀ ࡽ ࡢ 㑇 ఏ Ꮚ ࡘ ࠸ ࡚ ࠊ in situ hybridization ἲ Q-PCR ἲࢆ⏝࠸ࡓ☜ㄆᐇ㦂ࢆ ⾜ࡗࡓࠋ ࡼࡿྍ⬟ᛶࡶ࠶ࡿࡀࠊ⤖ㄽࢆᑟࡃࡣヨ⾜ᩘࡶࢹ࣮ ࢱࡢ㉁ࡶศ࡛ࡣ࡞࠸ࠋࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ࡢ⬻ࡢ୰ ࡽヱᙜࡍࡿᩘ༓ಶࡢ⚄⤒⣽⬊ࢆᡭసᴗ࡛ศྲྀࡋࠊ ࡑࡇࡽ RNA ࢆᢳฟࡍࡿ᪉ἲࢆྲྀࡗࡓࡓࡵࠊ༑ศ ࡞⣽⬊ᩘࢆ☜ಖࡍࡿࡇࡀ࡛ࡁࡎಙ㢗ᛶࡢ㧗࠸⤖ᯝ ࢆᚓࡽࢀ࡞ࡗࡓࠋࡲࡓグ᠈ᙧᡂࢆᢅ࠺௨ୖࠊከ㔞 ࡞ಶయࡽฟⓎࡍࡿࡇࡶ㞴ࡋࡃࠊ⤖ᯝⓗࡣᙜ ࡢ⚾ࡓࡕࡢᢏ⾡࡛ࡣ࡞ࡾᅔ㞴࡞ࣉࣟࢪ࢙ࢡࢺ࡛࠶ ࡗࡓࠋᮏᨭ࡛ࡣࡇࢀ௨ୖ⥅⥆ࡍࡿࡇࡀ࡛ࡁࡎṧ ᛕ࡛࠶ࡗࡓࠋ᭱᪂ࡢ⣽⬊ࣂ࣮ࢥ࣮ࢻࣛ࣋ࣝἲ࡞࡛ ࡇࡢㄢ㢟ࡶ࠺୍ᗘᣮᡓࡋ࡚ࡳࡓ࠸ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 pde1c ࡀ㛗ᮇグ᠈ᶵ⬟ࡍࡿࡇࡣ᪂ࡓ࡞▱ぢ ࠸࠼ࡿࠋ࢟ࣀࢥయ⚄⤒࡛Ⓨ⌧ࡍࡿ㑇ఏᏊࡢ㸦࠶ࡿ⛬ ᗘࡢ㸧ࣜࢫࢺࡀᚓࡽࢀࡓࡇ࡛グ᠈ࡢ⏕⌮Ꮫࢆ◊✲ ࡍࡿᇶ♏ⓗ࡞ࢹ࣮ࢱࢆᚓࡽࢀࡓࡇࡣࠊᮍࡔ᫂ࡽ ࡉࢀ࡚࠸࡞࠸ࢺࣛࣥࢫ࣑ࢵࢱ࣮ࡢྠᐃ࡞ᐤ ࡍࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ヱᙜ࡞ࡋࠋ cAMP ⤒㊰ࢆไᚚࡍࡿᅉᏊ - 425 - බເᨭㄢ㢟㸸2013 ᖺᗘࠊ2014 ᖺᗘ ༢᰾㈎㣗⣽⬊⩌ࡢศᶵᵓ㛵ࡍࡿ◊✲ ᇶ┙◊✲㸦㹀㸧 ⏣ᮧ ᬛᙪ࣭ᶓᕷ❧Ꮫ་Ꮫ㒊චᏛ 㸺⫼ᬒ࣭┠ⓗ㸼 ᑡᩘࡢ⣽⬊ࡽ㧗⢭ᗘࡢࢡ࣐ࣟࢳࣥචỿ㝆ࢤࣀ ࣒ ࢩ ࣮ ࢣ ࣥ ࢫ 㸦 chromatin immunoprecipitation sequencing, ChIP-seq㸧ࢆ⾜࡞࠺ࡇࡣ㸪ࢤࣀ࣒་ Ꮫ◊✲࠾࠸࡚ࡶ᭱ࡶ㔜せ࡞ㄢ㢟ࡢ୍ࡘᛮࢃࢀ㸪 ከࡃࡢ◊✲⪅ࡸᴗࡀࡇࢀྲྀࡾ⤌ࢇ࡛࠸ࡿࠋࡇࢀ ࡣ㸪ࡈࡃᚤ㔞ࡢ⣽⬊ࡋᚓࡽࢀ࡞࠸⏕యෆࡢ⣽⬊㸪 ≉࣐࢘ࢫࡢ⤌⧊ᖿ⣽⬊ࡸ๓㥑⣽⬊ࢆᢅ࠺ࡇࡢ㔜 せᛶ㉳ᅉࡍࡿࠋಶయࡢ⏕⌧㇟ࢆ▱ࡿୖ࡛㸪㑇ఏ ᏊḞᦆື≀ࢆᐜ᫆స〇࡛ࡁࡿ࣐࢘ࢫ⏕య⏤᮶⣽⬊ ࡛ࡢ㌿ᅉᏊࡸࣄࢫࢺࣥಟ㣭࡞ࡢືែࢆࢤࣀ࣒つ ᶍ࡛▱ࡿࡣᴟࡵ࡚㔜せ࡛࠶ࡾ㸪ࡑࡢ◊✲⤖ᯝࡢ⏕ ⌮Ꮫⓗព⩏ࡣ㸪⣽⬊ᰴࢆ⏝࠸ࡓ◊✲ࡃࡽ㐶 㧗࠸ࠋ ༢᰾㈎㣗⣽⬊㸦༢⌫࣭࣐ࢡࣟࣇ࣮ࢪ㸪ᶞ≧⣽⬊㸧 ࡣ⮬↛චཬࡧ⋓ᚓචࡢㄏᑟᚲ㡲ࡢච⣽⬊࡛ ࠶ࡾ㸪㐀⾑ᖿ⣽⬊ࡽ୰㛫ⓗ࡞๓㥑⣽⬊ẁ㝵ࢆ⤒࡚ ⏘⏕ࡉࢀࡿࠋ༢᰾㈎㣗⣽⬊ࡢศ࣭⏘⏕ࡢศᏊ࣓࢝ ࢽࢬ࣒ࢆ᫂ࡽࡍࡿࡓࡵࡣ㸪㐀⾑ᖿ⣽⬊ࡸ๓㥑 ⣽⬊࠾࠸࡚ࣄࢫࢺࣥಟ㣭ࡸ㌿ᅉᏊ⤖ྜࡢࢤࣀ࣒ つᶍ࡛ࡢᣲືࢆ ChIP-seq ࢆ⏝࠸࡚ゎᯒࡍࡿࡇࡀ ᴟࡵ࡚㔜せ࡛࠶ࡿࡀ㸪ࡇࢀࡽࡢ⣽⬊ࡣ⏕యෆᑡᩘ ࡋᏑᅾࡋ࡞࠸ࡓࡵゎᯒࡣᅔ㞴࡛࠶ࡗࡓࠋᮏᨭㄢ 㢟࡛ࡣࡲࡎ༢᰾㈎㣗⣽⬊๓㥑⣽⬊ࡢ ChIP-seq ゎᯒ ࢆ⾜࠺ࡓࡵ㸪ᑡᩘࡢ⣽⬊࡛ ChIP-seq ࢆ⾜࠺᪉ἲࢆ 㛤Ⓨࡍࡿࡇࢆ┠ᣦࡋࡓࠋࡉࡽᚤ㔞⣽⬊ࡼࡿ ChIP-seq ᢏ⾡ࢆ⏝࠸࡚㸪༢᰾㈎㣗⣽⬊ࡢ๓㥑⣽⬊ࡸ ࡑࢀ⏤᮶ࡍࡿᡂ⇍⣽⬊࠾ࡅࡿᵝࠎ࡞✀㢮ࡢࣄࢫ ࢺࣥಟ㣭ࡸ㌿ᅉᏊࡢ⤖ྜࡢࢤࣀ࣒つᶍゎᯒࢆ⾜࠺ ࡇࢆ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 2013 ᖺ ᗘ ࡣ ༢ ᰾ ㈎ 㣗 ⣽ ⬊ ࡢ ๓ 㥑 ⣽ ⬊ ࢆ ⏝ ࠸ ࡚ ChIP-seq ࢆ⾜࠺ࡓࡵ㸪⚾㐩ࡣ㌿ᅉᏊ IRF8 ࡸࣄ ࢫࢺࣥಟ㣭ࡢ ChIP ࡢ᮲௳᳨ウࢆ⾜ࡗࡓࠋࡑࡋ࡚ࢤ ࣀ࣒ᨭ࣭㕥ᮌ✨ඛ⏕ ChIP ࡋࡓ DNA ࢆḟୡ௦ࢩ ࣮ࢣࣥࢧ࣮ࡼࡿ㓄ิゎᯒࢆ⾜ࡗ࡚㡬࠸ࡓࠋࢧࣥࣉ ࣝㄪᩚࡣᅔ㞴ࢆకࡗࡓࡀ㸪10 ಶ௨ୗ࠸࠺ࡲ ࡛ࡢᶆ‽ⓗ࡞ࣉࣟࢺࢥ࣮ࣝࡢ࠾ࡼࡑ 10 ศࡢ 1 ࡢ⣽⬊ ᩘ࡛ ChIP-seq ࡀྍ⬟࡞ࡗࡓࠋᚤ㔞⣽⬊ࢆ⏝࠸ࡓ ChIP-seq ᢏ⾡ࡀ࠶ࡿ⛬ᗘ⌧ᐇࡢࡶࡢ࡞ࡗ࡚ࡁࡓ ࡇࡽ㸪2014 ᖺᗘࡣᢏ⾡ࢆࡉࡽ☜ᐇ࡞ࡶࡢ ࡍࡿࡶ㸪ูࡢ㌿ᅉᏊࡸࣄࢫࢺࣥಟ㣭ࡉࡽ 」ᩘࡢ༢᰾㈎㣗⣽⬊⩌๓㥑⣽⬊㸦㔝⏕ᆺ࣐࢘ࢫ㸪㌿ ᅉᏊ㑇ఏᏊḞᦆ࣐࢘ࢫ㸧ࡸ๓㥑⣽⬊ࡀ⏘⏕ࡍࡿᡂ ⇍㈎㣗⣽⬊ࢆྵࡵ࡚ໟᣓⓗ ChIP ࢆ⾜࠸㸪DNA 㓄 ิゎᯒࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ༢᰾㈎㣗⣽⬊ศ࠾ࡅࡿ๓㥑⣽⬊ࡽᡂ⇍⣽⬊ࡲ ࡛ࡢࣄࢫࢺࣥಟ㣭ࡢືែࡀ᫂ࡽ࡞ࡗࡓ㸦ㄽᩥᢞ ✏‽ഛ୰㸧 ࠋ≉㑇ఏᏊⓎ⌧࢚ࣥࣁࣥࢧ࣮ࣉ࣑ࣛ ࣥࢢ࣭άᛶࡢ㛵ಀࢆゎᯒࡍࡿ୰࡛㠀ᖖ⯆῝ ࠸ࢹ࣮ࢱࡀᚓࡽࢀࡓࠋࡲࡓ㌿ᅉᏊ IRF8 ࡸ PU.1 ࡢ ChIP-seq ࡶ⾜࠸㸪ࡇࢀࡽ㌿ᅉᏊࡀ༢᰾㈎㣗⣽⬊ࡢ ๓㥑⣽⬊࠾࠸࡚ࡢࡼ࠺࡞㑇ఏᏊ⤖ྜࡍࡿࡢ ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᚤ㔞⣽⬊ࡼࡿ ChIP-seq ࡀྍ⬟࡞ࡗࡓࡇࡽ㸪 ⏕యෆᏑᅾࡍࡿᵝࠎ࡞ᕼᑡ⣽⬊࡛ࡢ ChIP-seq ゎ ᯒࢆ㐍ࡵ࡚࠸ࡁࡓ࠸⪃࠼࡚࠸ࡿࠋ᭱㏆㸪㠀ᖖᑡ ᩘࡢ⣽⬊ࢆ⏝࠸ࡓ ChIP-seq ࢆྍ⬟ࡍࡿ᪉ἲࡀ࠸ ࡃࡘሗ࿌ࡉࢀ࡚࠸ࡿࡀ㸪ࡑࡢࢹ࣮ࢱ⚾㐩ࡢࢹ࣮ ࢱࡢẚ㍑ࢆ⾜ࡗࡓࡇࢁ㸪⚾㐩ࡢ࠺ࡀ᫂ࡽࢹ ࣮ࢱࡢ㉁ࡀ㧗࠸ࡇࡀࢃࡗ࡚࠸ࡿ㸦ᮍⓎ⾲ࢹ࣮ࢱ㸧ࠋ ච⣔ࡢ๓㥑⣽⬊ࡣᩘࡀᑡ࡞࠸ࡓࡵ㸪ᅇ㛤Ⓨࡋࡓ ᢏ⾡ࡀᚋ≉ࡁ࡞ຊࢆⓎࡍࡿ⪃࠼ࡽࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾᢞ✏‽ഛ୰࡛ࡍࠋ - 426 - බເᨭㄢ㢟㸸2015 ᖺᗘ ༢᰾㈎㣗⣽⬊⣔ࡢศ࠾ࡅࡿ㑇ఏᏊⓎ⌧ไᚚᶵᵓࡢໟᣓⓗゎ᫂ ᇶ┙◊✲㸦㹀㸧 ⏣ᮧ ᬛᙪ࣭ᶓᕷ❧Ꮫ་Ꮫ㒊චᏛ 㸺⫼ᬒ࣭┠ⓗ㸼 㐀⾑࠾ࡅࡿศࡣከẁ㝵⏕ࡌࡿࡀ㸪ྛศẁ㝵 ࡢ⣽⬊㞟ᅋࡣᚲࡎࡋࡶᆒ୍࡛ࡣ࡞ࡃ࠸ࡃࡘࡢள㞟 ᅋࡼࡗ࡚ᵓᡂࡉࢀ࡚࠸ࡿࡇࡀ㸪ḟࠎ᫂ࡽ ࡉࢀ࡚࠸ࡿࠋ≉ከศ⬟ࢆᣢࡘ㐀⾑ᖿ⣽⬊࣭๓㥑 ⣽⬊㸪≉ᐃࡢ⣽⬊✀ࡢࡳࡋ⏘⏕ࡋ࡞࠸⣽⬊ࡀ┦ ᙜᩘྵࡲࢀ࡚࠸ࡿࡇࡀࢃࡗ࡚ࡁ࡚࠸ࡿࠋࡓࡔࡋ ࡇࡢࠕ᪩ᮇࡢ㐠ỴᐃࠖࡢศᏊᶵᵓࡣࡃ࡛᫂࠶ ࡿࠋ ⚾㐩ࡣ⮬↛චࡸ⋓ᚓච࠾࠸࡚୰ᚰⓗᙺࢆ ᢸࡗ࡚࠸ࡿ༢᰾㈎㣗⣽⬊㸦༢⌫࣭࣐ࢡࣟࣇ࣮ࢪ㸪 ᶞ≧⣽⬊㸧ࡢศ࣭⏘⏕ࡢ࣓࢝ࢽࢬ࣒ࡘ࠸࡚◊✲ ࢆ⾜ࡗ࡚ࡁࡓࠋࡑࡢ୰࡛⚾㐩ࡣ㐀⾑ࡢୖὶ⨨ࡋ ከศ⬟ࢆᣢࡘ๓㥑⣽⬊ࡢ୍㒊ࡢศ⏬ࡼࡾࡶ㧗 ࠸༢᰾㈎㣗⣽⬊ศ⬟ࢆᣢࡘ㞟ᅋࡀᏑᅾࡍࡿࡇࢆ ぢฟࡋࡓࠋࡋࡋ࡞ࡀࡽ㸪ࡇࡢ๓㥑⣽⬊㞟ᅋࡶ༢᰾ ㈎㣗⣽⬊ࡢࡳศࡍࡿࢃࡅ࡛ࡣ࡞࠸ࡇࡽ㸪⚾ 㐩ࡣࡇࡢ⣽⬊㞟ᅋࡶࡲࡔᆒ୍࡛࠶ࡿ᥎ ࡋ࡚࠸ ࡿࠋࡑࡇ࡛ᅇࢤࣀ࣒ᨭ࣭㕥ᮌ✨ඛ⏕ࡢ༠ຊࢆᚓ ࡚㸪㐀⾑๓㥑⣽⬊ࡢ༢୍⣽⬊ RNA-seq ࢆ⾜࡞࠺ࡇ ࡛㸪ᇶ┙ B ◊✲ࡢᑐ㇟࡛࠶ࡿ༢᰾㈎㣗⣽⬊ࡢศ ࡼࡾ῝ࡃษࡾ㎸ࡳࠕ㐀⾑࠾ࡅࡿ᪩ᮇࢥ࣑ࢵࢺ࣓ࣥ ࢺࠖࡢศᏊᶵᵓ㏕ࡿࡇࡀ࡛ࡁࡿ⪃࠼࡚࠸ࡿࠋ ࡑࡋ࡚᭱⤊ⓗࡣࠕ⣽⬊㞟ᅋࡢᆒ୍ᛶࢆసࡾฟࡋ ࡚࠸ࡿ⤌ࡳࡑࡢ⏕≀Ꮫⓗព⩏ࠖࢆ᫂ࡽࡋࡓ ࠸ࠋ RNA-seq 㛵ࡍࡿᨭࢆ⾜ࡗ࡚㡬ࡃࠋලయⓗࡣ⚾ 㐩ࡀ༢୍⣽⬊ࡽྜᡂ࣭ቑᖜࡋࡓ cDNA ࡢ NGS ࣛ ࣈ࣮ࣛࣜࡢసᡂ DNA 㓄ิࡢỴᐃࢆ⾜ࡗ࡚㡬ࡃࠋ ࢧࣥࣉࣝᩘࡣྜィ 400 ࢧࣥࣉࣝ⛬ᗘࡢணᐃ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⌧ᅾ༢୍⣽⬊ࡽᚓࡽࢀࡓ cDNA ࡢ NGS ࣛࣈࣛ ࣮ࣜసᡂ DNA 㓄ิỴᐃࢆ⾜ࡗ࡚࠸ࡓࡔ࠸࡚࠸ࡿࠋ ࢹ࣮ࢱࡀᚓࡽࢀḟ➨㸪㏿ࡸゎᯒࢆ⾜࠸ࡓ࠸ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ⚾㐩ࡢ◊✲ᐊ࡛ࡣࡇࢀࡲ࡛༢୍⣽⬊ RNA-seq ࢆ ⾜ࡗࡓ⤒㦂ࡀ࡞ࡗࡓࠋ᭱ඛ➃ᢏ⾡ࡢᑟධࡣᵝࠎ ࡞㞀ቨࢆࡾ㉺࠼ࡿᚲせࡀ࠶ࡿࡀ㸪◊✲ㄢ㢟ࡀࢤࣀ ࣒ᨭ᥇ᢥࡉࢀࡓࡇࡀዎᶵ࡞ࡗ࡚㸪༢୍⣽⬊ RNA-seq ゎᯒࢆ⾜࠺ࡓࡵࡢ▱㆑ࡸᶵჾࢆಟᚓ࣭‽ഛ ࡍࡿࡇ⧅ࡀࡗࡓࠋࡇࢀࡽᚓࡽࢀࡿࢹ࣮ࢱࢆゎ ᯒࡋ㸪㏿ࡸⓎ⾲࡛ࡁࡿࡼ࠺ດࡵࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡲࡔࢹ࣮ࢱࡀᚓࡽࢀ࡚࠾ࡽࡎᨭࡢᡂᯝࡣ࡞࠸ࡀ㸪 ࡛ࡁࡿࡔࡅ᪩࠸ẁ㝵࡛ᏛⓎ⾲ࡸㄽᩥⓎ⾲ࡀ࡛ࡁࡿ ࡼ࠺ດຊࡋࡓ࠸⪃࠼࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ༢᰾㈎㣗⣽⬊ࡢศ࠾ࡅࡿ㑇ఏᏊⓎ⌧ไᚚ᪩ᮇ ࢥ࣑ࢵࢺ࣓ࣥࢺࡢศᏊ࣓࢝ࢽࢬ࣒ࢆ⌮ゎࡍࡿࡓࡵ㸪 ከศ⬟ࢆᣢࡘ㐀⾑๓㥑⣽⬊࠾ࡅࡿ༢୍⣽⬊ - 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Block of Cdk1-dependent poly(A) elongation of cyclin B mRNA in MI-arrested starfish oocytes is released by intracellular pH elevation upon spawning. Mol Reprod Dev 83 79-87 (2016) - 428 - බເᨭㄢ㢟㸸2013 ᖺᗘ ࢝ࣁࣛࣔࣜࡢࣔࢹࣝື≀ࢆຍ㏿ࡍࡿศ㔝ᶓ᩿◊✲㸸ᢏ⾡࣭ሗᇶ┙ᵓ⠏ᐇ㊶㐺⏝ ᇶ┙◊✲㸦㸿㸧 ༓ⴥ ぶᩥ࣭⟃ἼᏛ⏕⎔ቃ⣔ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢝ࣁࣛࣔࣜ㸦Cynops pyrrhogaster㸧ࡣࡑࡢࣘࢽ ࣮ࢡ࡞⣽⬊≉ᛶࡸ⣔⤫ⓗ⨨ࡽ⏕≀ᏛࡢṔྐṧ ࡿᩘከࡃࡢⓎぢ㈉⊩ࡋ࡚ࡁࡓᡃࡀᅜࢆ௦⾲ࡍࡿỈ ᳇ື≀㸦୧⏕㢮㸧࡛࠶ࡿࠋJapan Newt Research Community (JNRC)ࡣࠊࣔࣜ◊✲ࡢ᭱㔜せㄢ㢟ࡢ ୍ࡘ࡛࠶ࡿࠕ㈨※ࠖࠕࣔࢹࣝື≀ࠖࢆ᥎㐍ࡍ ࡃࠊつᶍᒇእ㣴Ṫᐇ㊶ࣞ࣋ࣝࡢࢺࣛࣥࢫࢪ࢙ ࢽࢵࢡ㸦TG㸧ࢆᐇ⌧ࡋࠊࡉࡽࡣࢤࣀ࣒ゎㄞࢆぢ ᤣ࠼ࡓ᪂つࣝࢦࣜࢬ࣒ࡢ㛤Ⓨࡶ╔ᡭࡋ࡚ࡁࡓࠋ ᡃࠎࡣ⌧ᅾࠊࡇࢀࡽࡢ㈨※࣭ᢏ⾡࣭ሗᇶ┙◊✲ࢆࠊ ࣔࣜࢆ⏝࠸ࡿᵝࠎ࡞◊✲ศ㔝ᒎ㛤ࡍࡃࠊTG ࢆ 㐺⏝ࡍࡿᛂ⏝ᢏ⾡ࡢ㛤Ⓨࠊ⏝౯್ࡢ㧗࠸㑇ఏᏊ ሗࡢ㞟㸦ࢹ࣮ࢱ࣮࣋ࢫᵓ⠏㸧ࢆ㐍ࡵ࡚࠸ࡿࠋ 㑇ఏᏊሗࡢ㞟άື࠾࠸࡚ࡣࠊ༓ⴥ┴㬞ᕝᕷ ⣔⤫ࣔࣜࢆ⏝࠸࡚ࠊࡍ࡛⢭ᕢ㸦㏻ᖖ㸧ࠊ㍺༸⟶ࠊ ༸ᕢࠊ⬇㸦16 ⣽⬊ᮇࠊ⚄⤒⬇ࠊᑿⱆ⬇㸧ࠊ⏕║⌫ 㸦⏕ึᮇࡢ⥙⭷Ỉᬗయ㸧ࡢ mRNA-seq ゎᯒࢆ⤊ ࡋࠊ⌧ᅾࠊ⢭ᕢ㸦࣏ࢺ࣮ࢩࢫ⌧㇟ࡀほᐹࡉࢀࡿ ≧ែࡢ⤌⧊㸧ࠊႥୖ⓶࣭㗟㰯ୖ⓶ࠊ⏕⫥㸦ึᮇ⏕ ⱆ㸧ࡢゎᯒࢆ㐍ࡵ࡚࠸ࡿࠋࡋࡋࠊࡇࢀࡔࡅ࡛ࡣ ሗࡀ༑ศ࡛࠶ࡿࠋࢥ࣑ࣗࢽࢸయࡢ◊✲ᒎ㛤ࢆ ᥎㐍ࡍࡿࡓࡵࡣࠊ࢝ࣁࣛࣔࣜⓎ⌧ࡍࡿ࡚ࡢ 㑇ఏᏊሗࢆ⥙⨶ࡋࡓ reference transcriptome ࢆᵓ ⠏ࡍࡿᚲせࡀ࠶ࡿࠋ ࡑࡇ࡛ࠊJNRC ࡣᮏᨭࡢࡶࠊ࢝ࣁࣛࣔࣜ reference transcriptome ࡢᵓ⠏ࢆ┠ᣦࡍࠋ2013 ᖺᗘ ࡣࠊ㑇ఏᏊሗࡢ⥙⨶ᛶࢆ☜ಖࡍࡿࡓࡵࠊṇᖖ࣭ ⏕⤌⧊㸦⬻ࠊ⥙⭷ࠊỈᬗయࠊᚰ⮚ࠊ⬨㧊㸧྾ჾ ⣔⤌⧊ࠊᾘჾ⣔⤌⧊ࡢ mRNA-seq ࢆ⾜࠺ࠋ 㸺ᨭࡢෆᐜ㸼 ⏕║⌫㸦⾡ᚋ 20 ᪥㸧 ࠊ⏕║⌫㸦⾡ᚋ 25 ᪥㸧 ࠊ ⏕║⌫㸦⾡ᚋ 30 ᪥㸧 ࠊ⏕║⌫㸦⾡ᚋ 50 ᪥㸧ࠊ⏕ ⬻ࠊ⏕ᚰ⮚ࠊỈᬗయࠊෆ⮚ჾᐁ㸦⫵ࠊ⭠ࠊ⮅⮚ࠊ ⫢⮚ࠊ⭈⮚㸧 ࡢ mRNA-seq 㸦Illumina HiSeq200-2500; 100bp paired-end read㸧ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 㞟ࡋࡓ࣮ࣜࢻሗࡽ⢭ᗘ⥙⨶ᛶࡢ㧗࠸ reference transcriptome ࡀ࡛ࡁࡿࡇ࡞ࡿࠋࡇࢀ ࡣࠊࣔࣜࡀࡶࡘ㑇ఏᏊࡢ✀㢮ࡸᩘࢆண ࡍࡿࡤ ࡾ࡛࡞ࡃࠊ࣐ࢵࣆࣥࢢࡼࡿ㑇ఏᏊࡢⓎ⌧ືែゎᯒ ࡸ࢟ࣕࣉࢳ࣮ࣕἲࡼࡿࢤࣀ࣒᩿∦㸦㌿ไᚚ㡿ᇦ ࡸࣥࢺࣟࣥ㸧ࡢྲྀᚓ࣭ゎᯒࠊࡉࡽࡣࢤࣀ࣒ᮏయ 㸦n=12㸹⣙ 20Gbp㸧ࡢゎᯒྥࡅ࡚᭷⏝࡞ࢶ࣮ࣝ ࡞ࡿ⪃࠼ࡽࢀࡿࠋᡃࠎࡣࡇࢀࢆ⏝ࡋ࡚ࠊ⤌⧊ࡈ ࡢ㌿⏘≀ண ࠊࣔࣜ≉␗ⓗ㑇ఏᏊண ࠊ Non-coding RNA ண ࢆ⾜࠸ࠊ2014-2015 ᖺᗘࡣ ࡇࢀࡽࡢሗࢆࡲࡵࡓࢹ࣮ࢱ࣮࣋ࢫࢆᵓ⠏࣭බ㛤 ࡍࡿࠋ ࡲࡓࠊᡃࠎࡣྠࡇࢀࡽࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ ሗࡢྛ◊✲⌧ሙࡢᐇ㊶㐺⏝ࢆ㐍ࡵ࡚࠾ࡾࠊ2014 ᖺᗘࡽࡇࡢྲྀࡾ⤌ࡳࢆᮏ᱁ࡉࡏࡿࠋࡍ࡛ࠊᑐ ㇟ࡍࡿ⏕⌧㇟㛵ࢃࡿೃ⿵㑇ఏᏊࡢ᥈⣴ࡸ㑇 ఏᏊࡢⓎ⌧ືែࡢẚ㍑ࠊ࠶ࡿ࠸ࡣ✀㛫ẚ㍑ࡼࡿ㐍 㐣⛬ࡢ⪃ᐹ⏝ࡉࢀጞࡵ࡚࠸ࡿࠋ࠼ࡤࠊ༓ⴥ 㸦⟃Ἴ㸧ࡣࠊ⏕≉␗ⓗ࡞ࣔࣜᅛ᭷㑇ఏᏊ㸦ࢱ ࣥࣃࢡ㉁ࢆࢥ࣮ࢻࡍࡿ᥎ᐃࡉࢀࡿ㸧ࡢೃ⿵ࢆ 14 ಶ ࡲ࡛⤠ࡾ㎸ࢇ࡛࠸ࡿࠋࡲࡓࠊࣔࣜࡀṇᖖ࡞Ỉᬗ యࢆ 20 ᅇ⧞ࡾ㏉ࡋ⏕࡛ࡁࡿࡇࢆド᫂ࡋࡘࡘ࠶ ࡿ㸦Tsonis࣭Ụཱྀࡢඹྠ◊✲㸧ࠋΏ㑓㸦ᒣᙧ㸧ࡣࠊ ཷ⢭㐣⛬㛵ࢃࡿ㑇ఏᏊẚ㍑ࡽయෆཷ⢭ᵝᘧࡢ㐍 ࡘ࠸࡚⪃ᐹࢆጞࡵ࡚࠸ࡿࠋ୰⏣㸦᪥ᮏ⋇་⏕ ⛉㸧ࡣᛶࣇ࢙ࣟࣔࣥ͂ࢯࢹࣇࣜࣥ̓ᑐࡍࡿႥཷ ᐜయࡢ㑇ఏᏊࢆ⤠ࡾ㎸ࢇ࡛࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Endo, Y. et al. Proceeding of the International Conference on Bioinformatics Models, Methods and Algorithms (BIOSTEC 2014), ISBN 978-989-758-012-3, 215-220 (2014). 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࢝ࣁࣛࣔࣜ reference transcriptome ࡢᵓ⠏༑ ศ࡞ሗࢆᚓࡿࡇࡀ࡛ࡁࡓࠋලయⓗࡣࠊᡃࠎ JNRC ഃࡀ⢭ᕢ㸦ṇᖖࠊ࣏ࢺ࣮ࢩࢫ㸧ࠊ㍺༸⟶ࠊ༸ ᕢࠊ⬇㸦16 ⣽⬊ᮇࠊ⚄⤒⬇ࠊᑿⱆ⬇㸧 ࠊ⏕║⌫㸦⾡ ᚋ 0-14 ᪥㸧 ࠊṇᖖ⫥ࠊ⏕⫥㸦⏕ⱆ㸧 ࠊႥୖ⓶࣭ࢪ ࣙࣅୖ⓶ࡽ⣙ 13.6 ൨࣮ࣜࢻࢆࠊࢤࣀ࣒ᨭഃࡀࠊ ⏕║⌫㸦⾡ᚋ 20 ᪥ࠊ25 ᪥ࠊ30 ᪥ࠊ50 ᪥㸧 ࠊ⏕ ⬻ࠊ⏕ᚰ⮚ࠊỈᬗయࠊෆ⮚ჾᐁ㸦⫵࣭⭠࣭⮅⮚࣭ ⫢⮚࣭⭈⮚㸧ࡽ⣙ 8.7 ൨࣮ࣜࢻࢆྲྀᚓࡋࡓࠋ - 429 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࢝ࣁࣛࣔࣜࡢࣔࢹࣝື≀ࢆຍ㏿ࡍࡿศ㔝ᶓ᩿◊✲㸸ᢏ⾡࣭ሗᇶ┙ᵓ⠏ᐇ㊶㐺⏝ ᇶ┙◊✲㸦㸿㸧 ༓ⴥ ぶᩥ࣭⟃ἼᏛ⏕⎔ቃ⣔ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢝ࣁࣛࣔࣜ㸦Cynops pyrrhogaster㸧ࡣࡑࡢࣘࢽ ࣮ࢡ࡞⣽⬊≉ᛶࡸ⣔⤫ⓗ⨨ࡽ⏕≀ᏛࡢṔྐṧ ࡿᩘከࡃࡢⓎぢ㈉⊩ࡋ࡚ࡁࡓᡃࡀᅜࢆ௦⾲ࡍࡿỈ ᳇ື≀㸦୧⏕㢮㸧࡛࠶ࡿࠋJapan Newt Research Community (JNRC)ࡣࠊࣔࣜ◊✲ࡢ᭱㔜せㄢ㢟ࡢ ୍ࡘ࡛࠶ࡿࠕ㈨※ࠖࠕࣔࢹࣝື≀ࠖࢆ᥎㐍ࡍ ࡃࠊつᶍᒇእ㣴Ṫᐇ㊶ࣞ࣋ࣝࡢࢺࣛࣥࢫࢪ࢙ ࢽࢵࢡ㸦TG㸧ࢆᐇ⌧ࡋࠊࡉࡽࡣࢤࣀ࣒ゎㄞࢆぢ ᤣ࠼ࡓ᪂つࣝࢦࣜࢬ࣒ࡢ㛤Ⓨࡶ╔ᡭࡋ࡚ࡁࡓࠋ ᡃࠎࡣ⌧ᅾࠊࡇࢀࡽࡢ㈨※࣭ᢏ⾡࣭ሗᇶ┙◊✲ࢆࠊ ࣔࣜࢆ⏝࠸ࡿᵝࠎ࡞◊✲ศ㔝ᒎ㛤ࡍࡃࠊTG ࢆ 㐺⏝ࡍࡿᛂ⏝ᢏ⾡ࡢ㛤Ⓨࠊ⏝౯್ࡢ㧗࠸㑇ఏᏊ ሗࡢ㞟㸦ࢹ࣮ࢱ࣮࣋ࢫᵓ⠏㸧ࢆ㐍ࡵ࡚࠸ࡿࠋ 㑇ఏᏊሗࡢ㞟άື࠾࠸࡚ࠊᡃࠎࡣ࢝ࣁࣛ ࣔ ࣜ ࡢ mRNA ሗ ࢆ ⥙ ⨶ ࡋ ࡓ reference transcriptome ࢹ࣮ࢱ࣮࣋ࢫࡢᵓ⠏ࢆ┠ᣦࡋ࡚࠸ࡿࠋ 2013 ᖺ ᗘ ᨭ ࡼ ࡿ ඹ ྠ ◊ ✲ ࡛ ࡣ ࠊ mRNA-seq 㸦Illumina HiSeq 2000-2500㸹100bp paired-end read㸧ࡼࡾࠊᡃࠎJNRC ഃࡀ⢭ᕢ㸦ṇᖖࠊ࣏ࢺ ࣮ࢩࢫ㸧ࠊ㍺༸⟶ࠊ༸ᕢࠊ⬇㸦16 ⣽⬊ᮇࠊ⚄⤒⬇ࠊ ᑿⱆ⬇㸧 ࠊ⏕║⌫㸦⾡ᚋ 0-14 ᪥㸧ࠊṇᖖ⫥ࠊ⏕⫥ 㸦⏕ⱆ㸧 ࠊႥୖ⓶࣭ࢪࣙࣅୖ⓶ࡽ⣙ 13.6 ൨࣮ࣜ ࢻࢆࠊࢤࣀ࣒ᨭഃࡀࠊ⏕║⌫㸦⾡ᚋ 20 ᪥ࠊ25 ᪥ࠊ30 ᪥ࠊ50 ᪥㸧ࠊ⏕⬻ࠊ⏕ᚰ⮚ࠊỈᬗయࠊෆ ⮚ჾᐁ㸦⫵࣭⭠࣭⮅⮚࣭⫢⮚࣭⭈⮚㸧ࡽ⣙ 8.7 ൨ ࣮ࣜࢻࢆྲྀᚓࡋࡓࠋᖺᗘࡣࠊࡇࢀࡽࡢሗࢆࡶ de novo assembly ࡼ ࡾ ࠕ ⤌ ⧊ ≉ ␗ ⓗ transcriptomeࠖ࠾ࡼࡧࠕ transcriptomeࠖࢆᵓ⠏ ࡍࡿࠋࡲࡓࠊࡇࢀࡽࢆཧ↷㓄ิࡋ࡚ࠊmRNA-seq/ ࣐ࢵࣆࣥࢢࡼࡿືⓗ transcriptome ゎᯒࢆヨࡳࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᗂ⏕ᚰ⮚㸦ኚែ๓㸧 ᡂయᚰ⮚㸦ኚែᚋ㸧 ࡢ mRNA-Seqࠊ ࠾ࡼࡧ༢㞳ࡋࡓ⥙⭷Ⰽ⣲ୖ⓶㸦RPE㸧⣽⬊ࣜࣉࣟ ࢢ࣒ࣛ୰ࡢ RPE ⣽⬊㸦RPE stem cell㸧ࡢቑᖜ cDNA ࡢ ḟ ୡ௦ ࢩ࣮ ࢣࣥ ࢫ㸦 ࠸ࡎ ࢀࡶ Illumina HiSeq 2000-2500㸹100bp paired-end read㸧 ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡚ ࡢ read ࢹ ࣮ ࢱ ࢆ ⏝ ࠸ ࡚ ࠊ Trinity 㸦 version 20131110㸧ࡼࡿ de novo assembly ࢆヨ⾜ࡋࡓ⤖ ᯝࠊ⥲ᩘ 697,670㸦N50: 2,380 b㸧ࡢࢥࣥࢸࢢࢆᚓ ࡿࡇࡀ࡛ࡁࡓࠋ⌧ᅾࠊሗᏛⓗゎᯒࡼࡾࡢ⬨ ᳝ື≀ࡢẚ㍑ࢆ㐍ࡵ࡚࠾ࡾࠊ㏆࠸ᑗ᮶ࠊ࢝ࣁࣛ ࣔࣜࡢࠕ transcriptomeࠖࡋ࡚Ꮫ⾡㞧ㄅሗ࿌ ࡍࡿィ⏬࡛࠶ࡿࠋྠ mRNA-seq ࢆ⾜ࡗࡓಶࠎࡢ ⤌⧊ࢧࣥࣉࣝ࠾࠸࡚ࡶ transcriptome ࢆᵓ⠏ࡋࡓࠋ ࡇࢀࡽࡢ⤌⧊≉␗ⓗ transcriptome ࡣࠊྛ◊✲⌧ሙ ࡢᚲせᛶࡶ࡙ࡁᵓ⠏ࡋࡓࡶࡢ࡛ࡶ࠶ࡾࠊゝࢃࡤ ಶࠎࡢࠕ⏕⌧㇟≉ࡋࡓ transcriptome࡛ࠖ࠶ࡿࠋ Reference transcriptome ࢆ⏝ࡋࡓ㑇ఏᏊⓎ⌧ ኚືゎᯒࡢඛ㥑ࡅࡋ࡚ࠊኚែ๓ᚋࡢᚰ⮚ࡽ mRNA-seq ࡼࡾࠊࡑࢀࡒࢀ⣙㸯൨࣮ࣜࢻᚓࡓࠋࡲ ࡓࠊࣜࣉࣟࢢ࣑ࣛࣥࢢ๓ᚋࡢ RPE ⣽⬊ࡑࢀࡒࢀ 100 ಶࡽ REPLI-g SensiPhi Polymerase ࢩࢫࢸ࣒ࢆ⏝ ࠸࡚ cDNA ࢆྜᡂ࣭ቑᖜࡋࠊࢩ࣮ࢣࣥࢫࡋࡓ㸦RPE ⣽⬊ࠊ4 ヨ⾜㸹RPE stem cellࠊ6 ヨ⾜㸹ྜィࠊ⣙ 5.6 ൨࣮ࣜࢻ㸧 ࠋ⌧ᅾࠊ⤌⧊≉␗ⓗࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢆཧ↷㓄ิࡋ࡚ࠊࡇࢀ ࡽࡢ࣐ࢵࣆࣥࢢゎᯒࢆ㐍ࡵ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᴗࡼࡗ࡚ᚓࡽࢀࢹ࣮ࢱࡣ࡚ࢥ࣑ࣗࢽࢸ ࣮ෆ࡛ඹ᭷ࡋ࡚࠸ࡿࠋ୍⯡ࡢ⏝⪅ᑐࡋ࡚ࡶ read ሗࡣ DDBJ/NCBI ࢆ㏻ࡌ࡚㸦2013 ᖺᗘᡂᯝሗ࿌㸧ࠊ ୍㒊ࡢ transcriptome ࡣ JNRC ࡀ⟶⌮ࡍࡿࢧࢺ IMORI (http://antler.is.utsunomiya-u.ac.jp/imori/) ࠾࠸࡚බ㛤ࡋࡓ㸦࡚ࡢ transcriptome ࡣ 2015 ᖺᗘ୰බ㛤ࡢணᐃ㸧 ࠋࡍ࡛ࢥ࣑ࣗࢽࢸ࣮࠾࠸ ࡚ࠊ㑇ఏᏊⓎ⌧ゎᯒࡸᶆⓗ࣭᪂ወ㑇ఏᏊࡢࢫࢡ࣮ࣜ ࢽࣥࢢࡢࡓࡵ⏝ࡀጞࡲࡗ࡚࠸ࡿࠋ⏕Ṫ⏕ࡢ ศ㔝࠾࠸࡚ࡣࠊㄽᩥࢆྜィ 3 ᮏⓎ⾲ࡋࡓࠋලయⓗ ࡣࠊ୧⏕㢮ࡢཷ⢭࣓࢝ࢽࢬ࣒ࡢከᵝࡢ㐍ⓗ㐨 ➽ࡢゎ᫂ࡸࠊᡂయࣔࣜ⥙⭷⏕ࡢᖿ࣓࢝ࢽࢬ࣒ 㸦ࣜࣉࣟࢢ࣑ࣛࣥࢢ㸧ࡢゎ᫂⧅ࡀࡗࡓࠋࡲࡓࠊⓎ ⏕ศ㔝࡛ࡣࠊTALEN/CRISPR ࡼࡿࢤࣀ࣒⦅㞟ᢏ ⾡ࡢᑟධホ౯ࠊኚែศ㔝࡛ࡣࠊኚែࡢࢲࢼ࣑ ࢵࢡ࡞⤌⧊ࣜࣔࢹࣜࣥࢢ㛵ࢃࡿᖿ⣽⬊ࡢືែゎᯒࠊ ⏕ศ㔝࡛ࡣࠊ⫥ࡢ⏕Ⓨ⌧ࡀቑຍࡍࡿࣔࣜ ᅛ᭷㑇ఏᏊࡢ༢㞳㸦⌧ᅾࡲ࡛ 2 ࡘࢆྠᐃ㸧ᶵ⬟ ゎᯒࠊ20 ᅇ⧞ࡾ㏉ࡋ⏕ࡉࡏࡓỈᬗయࡢရ㉁ホ౯ࠊ ⏕⌮ศ㔝࡛ࡣࠊỈ㝣୧⏝ࡢႥぬࢩࢫࢸ࣒ࡢゎᯒࡀ㐍 ࢇ࡛࠸ࡿࠋ ࡇࡢࡼ࠺ࠊᬑ㐢ᛶከᵝᛶࡢ୧᪉ࡢศ㔝࠾࠸ ࡚ࠊࡇࢀࡲ࡛ᅔ㞴࡛࠶ࡗࡓ㑇ఏᏊࣞ࣋ࣝࡢ◊✲ࡼ ࠺ࡸࡃ㊰ࡀ㛤ࢀࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Yokoe, M. et al., Int. J. Mol. Sci. 15, 15210-15224 (2014) Nakamura, K., et al., PLoS ONE 9, e109831 (2014) Islam, M.R. et al., Sci. Rep. 4, 6043 (2014) - 430 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࢝ࣁࣛࣔࣜࡢࣔࢹࣝື≀ࢆຍ㏿ࡍࡿศ㔝ᶓ᩿◊✲㸸ᢏ⾡࣭ሗᇶ┙ᵓ⠏ᐇ㊶㐺⏝ ᇶ┙◊✲㸦㸿㸧 ༓ⴥ ぶᩥ࣭⟃ἼᏛ⏕⎔ቃ⣔ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢝ࣁࣛࣔࣜ㸦Cynops pyrrhogaster㸧ࡣࡑࡢࣘࢽ ࣮ࢡ࡞⣽⬊≉ᛶࡸ⣔⤫ⓗ⨨ࡽ⏕≀ᏛࡢṔྐṧ ࡿᩘከࡃࡢⓎぢ㈉⊩ࡋ࡚ࡁࡓᡃࡀᅜࢆ௦⾲ࡍࡿỈ ᳇ື≀㸦୧⏕㢮㸧࡛࠶ࡿࠋJapan Newt Research Community (JNRC)ࡣࠊࣔࣜ◊✲ࡢ᭱㔜せㄢ㢟ࡢ ୍ࡘ࡛࠶ࡿࠕ㈨※ࠖࠕࣔࢹࣝື≀ࠖࢆ᥎㐍ࡍ ࡃࠊつᶍᒇእ㣴Ṫᐇ㊶ࣞ࣋ࣝࡢࢺࣛࣥࢫࢪ࢙ ࢽࢵࢡ㸦TG㸧ࢆᐇ⌧ࡋࠊࡉࡽࡣࢤࣀ࣒ゎㄞࢆぢ ᤣ࠼ࡓ᪂つࣝࢦࣜࢬ࣒ࡢ㛤Ⓨࡶ╔ᡭࡋ࡚ࡁࡓࠋ ᡃࠎࡣ⌧ᅾࠊࡇࢀࡽࡢ㈨※࣭ᢏ⾡࣭ሗᇶ┙◊✲ࢆࠊ ࣔࣜࢆ⏝࠸ࡿᵝࠎ࡞◊✲ศ㔝ᒎ㛤ࡍࡃࠊTG ࢆ 㐺⏝ࡍࡿᛂ⏝ᢏ⾡ࡢ㛤Ⓨࠊ⏝౯್ࡢ㧗࠸㑇ఏᏊ ሗࡢ㞟㸦ࢹ࣮ࢱ࣮࣋ࢫᵓ⠏㸧ࢆ㐍ࡵ࡚࠸ࡿࠋ 㑇ఏᏊሗࡢ㞟άື࠾࠸࡚ࠊᡃࠎࡣ 2013-2014 ᖺᗘࡢᨭࡼࡾࠊ࢝ࣁࣛࣔࣜࡢ⤌ ⧊≉␗ⓗ࠾ࡼࡧ transcriptome ࡢᵓ⠏ࢆ㛤ጞࡋࠊ ୍㒊ࡢሗࢆබ㛤ࡋࡓࠋࡲࡓࠊࡇࢀࡽࢆཧ↷㓄ิ ࡋ࡚ࠊ࣐ࢵࣆࣥࢢࡼࡿ㑇ఏᏊⓎ⌧ືែゎᯒࢆヨࡳ ࡿࡃࠊኚែ๓ᚋࡢᚰ⮚ࠊ࠾ࡼࡧࣜࣉࣟࢢ࣑ࣛࣥࢢ ๓ᚋࡢ⥙⭷Ⰽ⣲ୖ⓶㸦RPE㸧⣽⬊Ⓨ⌧ࡍࡿ mRNA ࡢࢩ࣮ࢣࣥࢫࢆ⾜ࡗࡓࠋᖺᗘࡣࠊࡇࢀࡽࡢࢩ࣮ࢣ ࣥࢫࢹ࣮ࢱࢆࡶ㑇ఏᏊࡢⓎ⌧ືែゎᯒࢆ⾜࠺ ࡶࠊᵝࠎ࡞ศ㔝ࡢ transcriptome ࡢᐇ㊶㐺⏝ 㸦㑇ఏᏊࡢ᥈⣴ࠊⓎ⌧ືែゎᯒࠊᶵ⬟ゎᯒ㸧ࢆࡉࡽ ᥎ࡋ㐍ࡵࡿࠋ 㸺ᨭࡢෆᐜ㸼 ➨ 3 ୡ௦➨ 7 ୡ௦ࡑࢀࡒࢀࡢ⢭ཎ⣽⬊ࡢ mRNA ࠾ ࡼࡧቑᖜ cDNAࠊ⏕ᚰ⮚ࡢ mRNAࠊPax6 㑇ఏᏊ ࢆࣀࢵࢡࢲ࢘ࣥࡋࡓ RPE ⣽⬊ࣜࣉࣟࢢ࣒ࣛ୰ࡢ RPE ⣽⬊ࡢቑᖜ cDNA ࡢࢩ࣮ࢣࣥࢫ㸦Illumina HiSeq 2000-2500㸹100bp paired-end read㸧 ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 2013-2015 ᖺᗘࡢᮏᨭࡢඹྠ◊✲ࡼࡾࠊ࢝ ࣁࣛࣔࣜࡢ⤌⧊≉␗ⓗ transcriptome ࢆ 21ࠊ࠾ࡼ ࡧ 1 ࡘࡢ transcriptome㸦ࢥࣥࢸࢢᩘ㸸694,138㸹 N50: 2,294 b 㸹 GC: 45.52% 㸧 ࢆ ᵓ ⠏ ࡋ ࡓ ࠋ transcriptome ࡣࠊde novo assembly ࡛ᚓࡽࢀࡓ ࢥࣥࢸࢢ㸦697,670㸧ࡽࣂࢡࢸࣜ⏤᮶ࡢࡶࡢࢆ 㝖࠸ࡓࡶࡢ࡛࠶ࡿࠋࡇࢀࡽࡢሗࡣࣀࢸ࣮ࢩࣙࣥ ⤖ ᯝ ࡶ ྵ ࡵ ࡚ ࠊ JNRC ࡀ ⟶ ⌮ ࡍ ࡿ ࢹ ࣮ ࢱ ࢧ ࢺ ‘MORI’㸦http://antler.is.utsunomiya-u.ac.jp/imori/) ࡽࢲ࣮࢘ࣥࣟࢻྍ⬟ࡍࡿ㸦2015 ᖺᗘෆ㸧 ࠋ ࡇࢀࡽࡢ㑇ఏᏊሗࡣࠊࣔࣜࢆ⏝࠸ࡿᵝࠎ࡞ศ 㔝࡛⏝ᐇ⦼ࡀୖࡀࡾࡘࡘ࠶ࡿࠋ࠼ࡤࠊ⏕ศ㔝 ࡛ࡣࠊ⫥⏕ࡢᵝᘧࡀᗂ⏕ᮇኚែᚋࡢᡂయ࡛ษࡾ ᭰ࢃࡿࡇࡸࠊ⫥⏕Ⓨ⌧ࡍࡿࣔࣜ⛉ᅛ᭷ࡢ 㑇ఏᏊࡀᏑᅾࡍࡿࡇࠊỈᬗయࡀຍ㱋ࡸ⧞ࡾ㏉ࡋ ⏕ᢠࡋ࡚ⱝ࠸≧ែࢆ⥔ᣢ࡛ࡁࡿࡇࠊ࡞ࢆ᫂ࡽ ࡋࡓࠋࡲࡓࠊࣔࣜ TTX ࡢ⏤᮶ࡢゎ᫂ࡸ de novo assemble ࣝࢦࣜࢬ࣒ࡢᨵⰋࡶ⧅ࡀࡗࡓࠋ 2014 ᖺᗘᚓࡓࢩ࣮ࢣࣥࢫࢹ࣮ࢱࡢ࣐ࢵࣆࣥ ࢢ࣭⤫ィゎᯒࡼࡾࠊRPE ⣽⬊ࡢࣜࣉࣟࢢ࣑ࣛࣥࢢ 㐣⛬࡛Ⓨ⌧㔞ࡀ᭷ពቑῶࡍࡿࢥࣥࢸࢢࢆ᫂ࡽ ࡋࡓࠋቑຍࡋࡓ 4,433 ῶᑡࡋࡓ 6,871 ࡣ᪤▱ࡢ 㑇ఏᏊࡢ┦ྠᛶࡀ࡞ࡗࡓࠋࡇࡢ୰ࡣ᭱ࡶ㢧ⴭ ࡞ቑຍࢆ♧ࡋࡓࢥࣥࢸࢢࡀྵࡲࢀ࡚࠸ࡓࠋ᪉ࠊ ┦ྠᛶࡀㄆࡵࡽࢀࡓࢥࣥࢸࢢࡢ࠺ࡕࠊቑຍࡋࡓ 5,407 ࡣࠊእയᛂ⟅ࡸୖ⓶-㛫ⴥ㌿ࠊ⣽⬊࿘ᮇࠊ ศከ⬟ᛶ㛵㐃ࡍࡿ㑇ఏᏊࡀከࡃྵࡲࢀ࡚࠸ࡓࠋ ῶᑡࡋࡓ 8,372 ࡣࠊRPE ⣽⬊ࡢ⏕⌮ᶵ⬟㛵㐃ࡍ ࡿ㑇ఏᏊࡀከࡃྵࡲࢀ࡚࠸ࡓࠋ ᖺᗘࡢᨭࡼࡗ࡚ᚓࡽࢀࡓࢩ࣮ࢣࣥࢫ⤖ᯝࡢ ゎᯒࡣ⌧ᅾ㐍⾜୰࡛࠶ࡿࠋࡇࢀࡽࡣࠊ⢭ཎ⣽⬊ࡢ ࣏ࢺ࣮ࢩࢫ㸦⢭Ꮚࡢရ㉁⟶⌮࣓࢝ࢽࢬ࣒ࡢ୍ࡘ㸧ࢆ ច㉳ࡍࡿศᏊ࣭⣽⬊࣓࢝ࢽࢬ࣒ࠊኚែ๓ᚋ࡛ኚࡍ ࡿᚰ⮚ࡢ⏕ᵝᘧࡑࢀࢆつᐃࡍࡿศᏊ࣭⣽⬊࣓࢝ ࢽࢬ࣒㸦2014 ᖺᗘࢩ࣮ࢣࣥࢫࢹ࣮ࢱࡶ⏝㸧ࠊ⥙⭷ ⏕ࡢศᏊ࣭⣽⬊࣓࢝ࢽࢬ࣒↔Ⅼࢆᙜ࡚࡚࠸ࡿࠋ ࡇࢀࡽࡢ◊✲ㄢ㢟ࡣࠊࡢࣔࢹࣝື≀࡛ࡣゎᯒᅔ㞴 ࡞⏕⌧㇟ࡢゎ᫂║ࡀ࠶ࡾࠊ་⒪ᛂ⏝ࡶ⧅ࡀ ࡿᬑ㐢ⓗၥ㢟࡛ࡶ࠶ࡿࡓࡵࠊ౯್ࡢ࠶ࡿ◊✲ᡂᯝࡀ ᮇᚅ࡛ࡁࡿࠋࡋࡶࠊࣔࣜࡢⅬࢆάࡋࡓゎീ ᗘࡢ㧗࠸ゎᯒ⤖ᯝࡣ࣐࢘ࢫ࣭ࣄࢺࢆ◊✲ࡍࡿ㝿ࡢࠕ≀ ᕪࠖࡋ࡚᭷⏝࡛࠶ࡿࠋ⌧ᅾ㐍⾜୰࡛࠶ࡿࡀࠊᡃࠎ ࡣࡇࢀࡽࡢ◊✲ࡼࡾᶆⓗ࣭᪂ወ㑇ఏᏊࢆࢫࢡ࣮ࣜ ࢽࣥࢢࡋࠊ⊂⮬ࡢ in vitro in vivo ࡢホ౯⣔ࡼࡾ ᮏ᱁ⓗ࡞ศᏊᶵ⬟ゎᯒ⧅ࡆࡓ࠸⪃࠼࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡀ࡞ࡅࢀࡤࠊ࢝ࣁࣛࣔࣜ transcriptome ࡢᵓ⠏ࡣᅔ㞴࡛࠶ࡗࡓࠋᮏᴗࡢ㛤ጞⅬ࡛ 200 ᮍ ‶ࡢ cDNA ሗࡋ࡞ࡗࡓࡇࡢື≀ࡢ◊✲࣏ࢸࣥ ࢩࣕࣝࡣ㣕㌍ⓗྥୖࡋࡓࠋ≉ࠊ᭱ඛ➃࣑࢜ࢵࢡ ࢫᢏ⾡ࡢ㐺⏝ࡀྍ⬟࡞ࡗ࡚ࡁࡓࡇࡣࡁ࠸ࠋ 㸦 ⏕ศ㔝࡛ࡣࠊ࣓࢝ࢽࢬ࣒ゎ᫂་⒪ᛂ⏝ࡢ㐨➽ࡀ ぢ࠼࡚ࡁࡓゝࡗ࡚ࡶࡼ࠸㸧ࠋ⌧ᅾ㐍ࡵ࡚࠸ࡿᶆⓗ࣭ ᪂ወ㑇ఏᏊࡢࢫࢡ࣮ࣜࢽࣥࢢᶵ⬟ゎᯒࢆࢽࢩ ࢸࣈࡋ࡚ࠊࢥ࣑ࣗࢽࢸ࣮యࡢ◊✲ࢫࢱࣝ ࡀࡁࡃኚㇺࡍࡿࡇࡣ࠸࡞࠸ࠋ transcriptome ࢆᚓࡓࡇ࡛ࠊࣔࣜࢤࣀ࣒㸦n=12, ~20 Gbp㸧ࡢゎ ㄞྥࡅ୍࡚Ṍ㋃ࡳฟࡏࡓ⪃࠼࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Inami, W. et al., Zool. Sci. (2016) in press Sousounis, K. et al., eLife 2015;10.7554/eLife.09594. (2015) Kudo, Y. et al., Toxicon 101, 101-105 (2015) Endo, Y. et al., IPSJ Transactions on Bioinformatics 8, 2-8 (2015) Casco-Robles, M.M. et al., Transgenic Res. 24, 463-473 (2015) - 431 - බເᨭㄢ㢟㸸2012 ᖺᗘ Ⓨ⏕㐣⛬࡛⣽⬊㐠ࢆつᐃࡍࡿ㌿ᅉᏊࡢ⏕ᾭࢃࡓࡿⓎ⌧ไᚚᶵᵓ ⱝᡭ◊✲㸦㹀㸧 ሯཎ 㐩ஓ࣭ᮾிᏛᏛ㝔⌮Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⬨ ᳝ ື ≀ ࠾ ࠸ ࡚ ࠊ DNA ప ࣓ ࢳ ࣝ 㡿 ᇦ 㸦Hypomethylated domain; HMD㸧ࡣ㑇ఏᏊࡢࣉࣟ ࣮ࣔࢱ࣮㡿ᇦᏑᅾࡋࠊ㑇ఏᏊࡢ㌿άᛶാࡃ ⪃࠼ࡽࢀ࡚ࡁࡓࠋ㏆ᖺࠊES ⣽⬊࡞ከ⬟ᛶࢆ᭷ࡍ ࡿ⣽⬊࠾࠸࡚ࠊᢚไᆺࣄࢫࢺࣥಟ㣭࡛࠶ࡿ H3K27me3 ࠾ ࡼ ࡧ ά ᛶ ᆺ ࣄ ࢫ ࢺ ࣥ ಟ 㣭 ࡛ ࠶ ࡿ H3K4me2 ࡢ୧⪅ࢆྵࡴ HMD㸦K27HMD㸧ࡀࠊⓎ⏕࣭ ศ㔜せ࡞ᙺࢆᯝࡓࡍ㑇ఏᏊୖከࡃᏑᅾࡍࡿ ࡇࡀ᫂ࡽ࡞ࡾࠊ㑇ఏᏊⓎ⌧ไᚚ࠾ࡅࡿ K27HMD ࡢᙺࡀὀ┠ࡉࢀ࡚࠸ࡿࠋᡃࠎࡣࠊ࣓ࢲ࢝ ึᮇ⬇ࢆ⏝࠸ࡓゎᯒࡽࠊK27HMD ࡣ 4kb ௨ୖࡢ large K27HMD ࠾ࡼࡧ 4kb ௨ୗࡢ small K27HMD ࠸ ࠺␗࡞ࡿ≉ᚩࢆᣢࡘࢧࣈࢱࣉࡀᏑᅾࡋࠊ๓⪅ࡣ⣽ ⬊ࡢ㐠ࢆつᐃࡍࡿ㌿ᅉᏊࢆࠊᚋ⪅ࡣⓎ⏕㔜せ ࡞ࢩࢢࢼࣝศᏊࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࢆከࡃྵࡴࡇ ࢆ᫂ࡽࡋࡓࠋ⯆῝࠸ࡇࠊK27HMD ࡢࢧ ࢬ H3K27me3 ࣞ࣋ࣝࡣṇࡢ┦㛵ࡀ࠶ࡾࠊlarge K27HMD ྵࡲࢀࡿ㑇ఏᏊࡣࡼࡾᙉࡃⓎ⌧ࡀᢚไࡉ ࢀ࡚࠸ࡿࡇࡀ♧၀ࡉࢀࡓࠋࡲࡓࠊศࡋࡓ⣽⬊ ࠾࠸୍࡚㒊ࡢ K27HMD ࡣࢧࢬࡀ⦰ᑠࡋ࡚࠾ࡾࠊࡑ ࡢࡼ࠺࡞ HMD ྵࡲࢀࡿ㑇ఏᏊࡣⓎ⌧ࡀୖ᪼ࡋ࡚ ࠸ࡿࡇࡶぢฟࡋࡓࠋࡇࢀࡽࡢ▱ぢࡽࠊK27HMD ࡣࢧࢬ౫Ꮡⓗ㑇ఏᏊⓎ⌧ࡢไᚚ≧ែࡀኚࡍࡿ ࠸࠺ྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋࡉࡽࠊ㏆ᖺங㢮ࡢ ᡂయ࠾࠸࡚ DNA ࡢࣄࢻࣟ࢟ࢩ࣓ࢳࣝ㸦5hmC㸧 㑇ఏᏊⓎ⌧ࣞ࣋ࣝṇࡢ┦㛵ࡀ࠶ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡋࡓࡀࡗ࡚ࠊᡃࠎࡢぢ࠸ࡔࡋࡓ K27HMD ࡢ⦰ᑠ 5hmC ࡀ㛵ࡍࡿྍ⬟ᛶࡀ࠶ࡿࠋ ࡑࡇ࡛ᡃࠎ ࡣࠊࡇࢀࡽࡢྍ⬟ᛶࢆ᳨ドࡍࡿࡓࡵ࣓ࢲ࢝ࢆ⏝࠸࡚ RNA-seq࣭ ChIP-seq࣭DNA ࣄࢻࣟ࢟ࢩ࣓ࢳࣝゎᯒ ࢆ⾜ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 ձ RNA-seqゎᯒ㸦࣓ࢲ࢝ᡂ㨶⫢⮚ࠊ1⣽⬊ᮇࠊ⬊⬇ ᮇ㸦๓ᮇ࣭ᚋᮇ㸧ࠊ๓ᮇཎ⭠⬇ᮇ㸧 ղ ChIP-seqゎᯒ㸦࣓ࢲ࢝ᡂ㨶⫢⮚࠾ࡅࡿH3K4me2 ࠾ࡼࡧH3K27me3㸧 ճ ࣁࢻࣟ࢟ࢩ࣓ࢳࣝゎᯒ㸦࣓ࢲ࢝ᡂ㨶⫢⮚ࠊ⫼ ഃ➽⠇ࠊ⭡ഃ➽⠇㸧 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ձ1⣽⬊ᮇࠊ⬊⬇ᮇࠊ๓ᮇཎ⭠⬇ᮇࡢ⣽⬊ࢆ⏝࠸࡚ RNA-seqゎᯒࢆ⾜࠸ࠊlarge K27HMDsmall K27HMD ྵࡲࢀࡿ㑇ఏᏊࡢⓎ⌧ࣞ࣋ࣝࢆẚ㍑ࡋࡓࡇࢁࠊ1 ⣽⬊ᮇ࠾࠸࡚ࡣ᭷Ⅽlarge K27HMDྵࡲࢀࡿ㑇 ఏᏊࡢⓎ⌧ࣞ࣋ࣝࡀపࡗࡓࠋ୍᪉ࠊ⬊⬇ᮇ࠾ࡼࡧ ཎ⭠⬇ᮇ࠾࠸࡚ࡣྠᵝࡢഴྥࡀぢࡽࢀࡓࡶࡢࡢ⤫ ィⓗࡣ᭷Ⅽ࡞ᕪࢆ᳨ฟฟ᮶࡞ࡗࡓࠋࡇࢀࡣࠊ࣓ ࢲ࢝ࢆྵࡴ㨶㢮ࡣẕᛶRNAࡢᙳ㡪ࢆᙉࡃཷࡅࡿࡓࡵ ࠊึᮇ⬇࠾ࡅࡿRNA-seqࡢ⤖ᯝࡣṇ☜࡞㌿㔞ࢆ ᫎࡋ࡞࠸ࡇࠊ⬊⬇ᮇࡸཎ⭠⬇ᮇࡣzygotic gene activation࠸࠺ࢤࣀ࣒࣡ࢻ㌿ࡀᙉຊάᛶ ࡍࡿᮇࡢ┤ᚋ࡛࠶ࡿࡓࡵRNA-seqࡢ⤖ᯝࡀ㑇ఏᏊ ࡢ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞≧ែࢆᚲࡎࡋࡶᫎࡋ࡞࠸ ࡇࡀཎᅉ࡛࠶ࡿ᥎ ࡉࢀࡿࠋ ղᡂ㨶⫢⮚࠾࠸୍࡚㒊ࡢ K27HMD ࡣࢧࢬࡢ⦰ᑠ ࡀㄆࡵࡽࢀࡿࡇࡽࠊRNA-seq ࡼࡾ㑇ఏᏊⓎ⌧ ࣞ࣋ࣝࡢ㛵㐃ࢆㄪࡓࠋࡑࡢ⤖ᯝࠊ⦰ᑠࡀぢࡽࢀ ࡓ K27HMD ࡛ࡣ H3K27me3 ࡢࣞ࣋ࣝࡶపୗࡋ࡚࠾ࡾࠊ 㑇ఏᏊⓎ⌧ࡣୖ᪼ࡋ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ᡃࠎࡣ➽⠇࠾࠸࡚ࡶྠᵝࡢ⤖ᯝࢆᚓ࡚࠾ࡾࠊᚋᮇ ࢫࢸ࣮ࢪ⏕ࡌࡿ K27HMD ࡢ⦰ᑠక࠺㑇ఏᏊⓎ⌧ ࣞ࣋ࣝࡢஹ㐍ࡀከࡃࡢ⤌⧊࡛ᬑ㐢ⓗぢࡽࢀࡿ⌧㇟ ࡛࠶ࡿࡇࢆ♧၀ࡋࡓࠋࡉࡽࠊᡂ㨶⫢⮚࠾࠸࡚ ⦰ᑠࡀㄆࡵࡽࢀ࡞࠸ K27HMD ╔┠ࡋࠊࡑࡢࢧࢬ 㑇 ఏ Ꮚ Ⓨ ⌧ ࣞ ࣋ ࣝ ࢆ ẚ ㍑ ࡋ ࡓ ࡇ ࢁ ࠊ large K27HMD ࠾࠸࡚㑇ఏᏊࡢⓎ⌧ࡀᴟࡵ࡚పࡃ࡞ࡗ࡚ ࠾ࡾࠊᙉᅛⓎ⌧ࡀᢚไࡉࢀ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡇࡢ⤖ᯝࡣࠊึᮇ⬇࡛♧၀ࡉࢀࡓ K27HMD ࡢࢧࢬ౫Ꮡⓗ࡞㑇ఏᏊᢚไᶵ⬟ࡘ࠸࡚ࠊᡂయ ࠾࠸࡚ࡶྠᵝࡢᶵ⬟᭷ࡍࡿྍ⬟ᛶࢆ♧ࡋ࡚࠸ࡿࠋ ճhydroxymethyl collector ࢆ⏝࠸࡚ᡂ㨶⫢⮚࣭⫼ഃ➽ ⠇࣭⭡ഃ➽⠇࠾ࡅࡿ 5hmC ゎᯒࢆ⾜࠸ࠊᚋᮇࢫࢸ ࣮ࢪ⏕ࡌࡿ DNA ࣓ࢳࣝࡢ୍㒊ࡣ 5hmC ࡛࠶ࡿࡇ ࢆぢฟࡋࡓࠋࡋࡋ࡞ࡀࡽࠊࢤࣀ࣒࣡ࢻ࡞ศᕸ ࢆヲ⣽ゎᯒࡋࡓࡇࢁࠊᚋᮇࢫࢸ࣮ࢪ࡛⏕ࡌࡓ DNA ࣓ࢳࣝࡼࡗ࡚ K27HMD ࡢ⦰ᑠࡢ㉳ࡁࡿࢤ ࣀ࣒㡿ᇦࡑࢀ௨እࡢ࣓ࢳࣝࡉࢀࡓ㡿ᇦࡢ㛫 5hmC ࣞ࣋ࣝᕪࡣぢࡽࢀ࡞ࡗࡓࠋࡋࡓࡀࡗ࡚ࠊᚋ ᮇࢫࢸ࣮ࢪ࠾࠸࡚ K27HMD 㡿ᇦ⏕ࡌࡿ DNA ࣓ ࢳࣝࡣ㑇ఏᏊⓎ⌧ࡢୖ᪼ࢆక࠺ࡀࠊ≉␗ⓗ࡞ࣄࢻ ࣟ࢟ࢩࡣぢཷࡅࡽࢀ࡞࠸ࡇࡀ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ࡼࡾࠊᡂయ࠾ࡅࡿ K27HMD ࡢ⦰ᑠ㑇ఏᏊ Ⓨ⌧ṇࡢ┦㛵㛵ಀࡀࠊᵝࠎ࡞⤌⧊࠾࠸࡚ᬑ㐢ⓗ ぢࡽࢀࡿ⌧㇟࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋࢢ࣮ࣝࣉ ࡢሗ࿌ࡼࡾ DNA ࣓ࢳࣝ H3K27me3 ࡢᣕᢠ㛵 ಀࡀ♧၀ࡉࢀ࡚࠸ࡿࡇࠊK27HMD ࡢ⦰ᑠࢆᘬࡁ㉳ ࡇࡍ DNA ࣓ࢳࣝࡀ⏕ࡌ࡚ࡶࣉ࣮ࣟࣔࢱ࣮㡿ᇦࡣ ప࣓ࢳࣝ≧ែࡀಖࡓࢀࡿࡇ࡞ࡽࠊᚋᮇࢫࢸ ࣮ࢪ⏕ࡌࡿ DNA ࣓ࢳࣝࡣ H3K27me3 ࢆᢚไࡍ ࡿࡇ࡛㌿άᛶ≧ែࡢ⥔ᣢᐤࡍࡿྍ⬟ᛶࡀ ⪃࠼ࡽࢀࡿࠋK27HMD ࡣ⣽⬊ࡢ㐠ࢆไᚚࡍࡿ㑇 ఏᏊࡀከࡃྵࡲࢀࡿࡇࡽࠊK27HMD ࡢ⦰ᑠࡣࡑ ࢀࡽࡢ㑇ఏᏊࡢⓎ⌧⥔ᣢࢆࡋ࡚⣽⬊࠾ࡼࡧ⤌⧊ࡢ ᜏᖖᛶ⥔ᣢാࡃ࠸࠺᪂ࡓ࡞௬ㄝࢆᚓࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Nakamura, R. et al., Development, 141, 2568-1580 (2014) - 432 - බເᨭㄢ㢟㸸2013 ᖺᗘ άᛶᆺ㌿⛣ᅉᏊࡀࢿࡢ࢚ࣆࢤࣀ࣒࠾ࡼࡰࡍຠᯝࡢゎ᫂⫱✀ࡢᛂ⏝ ᇶ┙◊✲㸦㹀㸧 ⠏ᒣ ᣅྖ࣭ி㒔ᏛᏛ㝔㎰Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ㏆ᖺࠊሷᇶ㓄ิከᆺᇶ࡙ࡃࣞࣝࡢࡳ࡞ࡽࡎࠊ ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ಟ㣭ࡢ㐪࠸ᇶ࡙ࡃ࢚ࣆࣞ ࣝࢆ⏝ࡋࡓ⫱✀ἲࡢ㛤Ⓨࡀ㐍ࡵࡽࢀ࡚࠸ࡿࠋࢃࢀ ࢃࢀࡣࠊỈ✄ရ✀ࠕ㖟ᆓࠖࡢȚ⥺↷ᑕᚋ௦࡛ᚓࡽ ࢀࡓ⣽⢏✺↛ኚ␗⣔⤫ IM294 ࡢ᫆ኚᛶࡢゎᯒ㐣⛬ ࠾࠸࡚ࠊື᳜≀ࢆ㏻ࡌ࡚ึࡵ࡚㌿⛣άᛶࢆࡶࡘ MITE (miniature inverted-repeat transposable element)ࠊmPing ࢆྠᐃࡋࡓࠋࢃࢀࢃࢀࡢࡇࢀࡲ࡛ ࡢ◊✲ࡽࠊmPing ࡣ㏆ഐࡢ㑇ఏᏊࡢⓎ⌧ࢫࢺࣞ ࢫᛂ⟅ᛶࢆࡍࡿࡇࠊmPing ࡢ࣓ࢳࣝ≧ែࡀ ࢫࢺࣞࢫࡼࡗ࡚ኚࡍࡿሙྜࡀ࠶ࡿࡇࠊ࠾ࡼࡧ mPing ࡣᐟࡢ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ไᚚࡢᙳ㡪ࢆ ཷࡅࡃ࠸ࡇࡀ᫂ࡽ࡞ࡗ࡚࠸ࡿࠋᮏ◊✲ࡣࠊ mPing ࡀᐟࡢ࢚ࣆࢤࣀ࣒࠾ࡼࡰࡍຠᯝࢆゎᯒࡍ ࡿࡶࠊmPing ࡢ㌿⛣ᐟࡢ࢚ࣆࢪ࢙ࢿࢸ ࢵࢡ࡞ไᚚࡢ㛵ಀࢆゎ᫂ࡍࡿࡇ࡛ࠊ㌿⛣ᅉᏊࢆ⏝ ࠸ࡓ࢚ࣆࢤࣀ࣒⫱✀ࡢᇶ┙ࢆᵓ⠏ࡋࡼ࠺ࡍࡿࡶࡢ ࡛࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᮏᨭ࡛ࡣࠊmPing ࡀᐟࡢ࢚ࣆࢤࣀ࣒≧ែ࠾ ࡼࡰࡍຠᯝࢆ᫂ࡽࡍࡿࡓࡵࠊḟୡ௦ࢩ࣮ࢣࣥ ࢧ࣮ࢆ⏝࠸࡚ mPing ࡢෆ㒊࠾ࡼࡧ㏆ഐ㓄ิࡢ࣓ࢳࣝ ⛬ᗘࢆつᶍゎᯒࢆヨࡳࡓࠋࡇࢀࡲ࡛ࡢ◊✲ࡽࠊ 㖟ᆓ࠾࠸࡚ mPing ࡣ 1,000 ࢥࣆ࣮௨ୖᏑᅾࡋࠊ ࡑࡢ࣓ࢳࣝ⛬ᗘࡣࢥࣆ࣮㛫࡛␗࡞ࡿࡇࡀ᫂ࡽ ࡞ࡗ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚ࠊmPing ࡢ࣓ࢳࣝࢆゎ ᯒࡍࡿ㝿ࡣࠊ1,000 ࢥࣆ࣮௨ୖࡢᤄධ⨨ࢆ᫂☜ ༊ูࡍࡿᚲせࡀ࠶ࡿࠋࡋࡓࡀࡗ࡚ࠊᮏᨭ࡛ࡣࠊࡲ ࡎࠊ㖟ᆓࡢⴥ㌟ࡽᢳฟࡋࡓࢤࣀ࣒ DNA ࢆ㗪ᆺ ࡋ࡚ࠊmPing ࡢࣉࣛࢫ㙐ࡢ 4 ⟠ᡤタィࡋࡓࣉࣛ ࣐ ࣮ ࢆ ⏝ ࠸ ࡚ Post-bisulfite Adaptor Tagging (PBAT)ἲ࡛ mPing 㓄ิ≉␗ⓗ࡞ࣛࣈ࣮ࣛࣜࢆస ᡂࡋࡓࠋࡑࡋ࡚ࠊHiSeq ࡼࡿࢩ࣮ࢣࣥࢫࢆ⾜ࡗࡓ ᚋࠊࣂࢧࣝࣇࢺࢩ࣮ࢣࣥࢫゎᯒࢶ࣮ࣝ Bismark ࡼࡗ࡚ mPing 㓄ิ࠾ࡼࡧࢿࢤࣀ࣒ ࣐ࢵࣆࣥࢢࡋࡓࠋ 58%࠾ࡼࡧ 31%࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋᮏ◊ ✲⤖ᯝࢆᇶࠊࡇࢀࡲ࡛ࡢ◊✲࡛᫂ࡽ࡞ࡗ࡚࠸ ࡿ 16 ⟠ᡤࡢ mPing ᤄධ⨨ࡢ࣓ࢳࣝ⛬ᗘࢆẚ㍑ ゎᯒࡋࡓࡇࢁࠊάᛶᆺ mPing(㌿⛣ࡀࡳࡽࢀࡿࢥࣆ ࣮)࡛ࡣࢇ࡚ࡢࢩࢺࢩࣥࡢ࣓ࢳࣝࡀపୗ ࡋ࡚࠸ࡿࡢᑐࡋ࡚ࠊάᛶᆺ mPing (㌿⛣ࡀࡳࡽ ࢀ࡞࠸ࢥࣆ࣮)࡛ࡣ࣓ࢳࣝࡢపୗࡣࢇぢࡽ ࢀ࡞ࡗࡓࠋࡇࡢࡇࡽࠊ࣓ࢳࣝ⛬ᗘࡢῶᑡࡀ mPing ࢆάᛶࡍࡿせᅉ࡛࠶ࡿࡇࡀ☜ࡵࡽࢀࡓࠋ ࡋࡋࠊάᛶ mPing ࡢ୰ࡶ࣓ࢳࣝ⛬ᗘࡢప ࠸ࡶࡢࡀྵࡲࢀ࡚࠸ࡓࡇࡽࠊmPing ࡣ DNA ࣓ ࢳࣝࡢࡳࡼࡗ࡚άࡉࢀ࡞࠸ࡢ࡛ࡣ࡞࠸ ⪃࠼ࡽࢀࡓࠋ୍᪉ࠊάᛶᆺ mPing ࡛ࡣࠊࢧࢺ≉␗ ⓗ࣓ࢳࣝࡢᖜ࡞పୗࡀࡳࡽࢀࡓࡇࡽࠊ≉ ᐃࡢࢩࢺࢩࣥṧᇶࡀ㌿⛣άᛶ㛵ࡋ࡚࠸ࡿྍ⬟ᛶ ࡀ࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡢ⤖ᯝࠊmPing ࡢ㌿⛣ࡣࠊ㓄ิ୰ࡢ≉ᐃࡢ ࢩࢺࢩࣥṧᇶࡢ࣓ࢳࣝࡼࡗ࡚ไᚚࡉࢀ࡚࠸ࡿྍ ⬟ᛶࡀ♧၀ࡉࢀࡓࠋࡋࡋࠊ2013 ᖺᗘࡣࠊࣛࣈࣛ ࣮ࣜࡢㄪᩚࡀ༑ศ࡛࠶ࡗࡓࡓࡵࠊᤄධ⨨ࢆỴᐃ ࡛ࡁࡓࢥࣆ࣮ࡀᑡ࡞ࡃࠊࢥࣆ࣮㛫ࡢ࣓ࢳࣝ⛬ᗘࢆ ༑ศẚ㍑࣭ゎᯒࡍࡿࡇࡀ࡛ࡁ࡞ࡗࡓࠋ⌧ᅾࠊ PBAT ἲࢆᨵⰋࡋࠊmPing 㓄ิ≉␗ⓗ࡞ࣛࣈࣛࣜ ࣮ࡢᵓ⠏ࢆヨࡳ࡚࠸ࡿࠋᚋࡣࠊᚓࡽࢀࡓࣛࣈ ࣮ࣛࣜࢆ⏝࠸࡚ࠊୖグྠᵝࡢ᪉ἲ࡛ゎᯒࢆ⾜࠸ࠊ ࢥࣆ࣮㛫ࡢ࣓ࢳࣝ≧ែࡢ㐪࠸ࢆ᫂ࡽࡋࡓ࠸ ⪃࠼࡚࠸ࡿࠋࡇࢀࡽࡢ⤖ᯝࡼࡗ࡚ࠊ㌿⛣ᅉᏊࢆ⏝ ࠸ࡓ࢚ࣆࢤࣀ࣒⫱✀ࡢᇶ┙ࡢᵓ⠏ࡢࡳ࡞ࡽࡎࠊ㌿⛣ ᅉᏊࡀࢤࣀ࣒୰࡛ࡢࡼ࠺ࡋ࡚ቑṪࡋࠊࡢࡼ࠺ ࡋ࡚㐍ࢆ≌ᘬࡍࡿࢆ⌮ゎࡍࡿ㔜せ࡞ᡭࡀࡾ ࡀᚓࡽࢀࡿᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ヱᙜ࡞ࡋࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᮏᨭࡢ⤖ᯝࠊ⥲࣮ࣜࢻᩘ 146,467,644 ࡢࢩ࣮ࢣ ࣥࢫࢹ࣮ࢱࢆᚓࡓࠋࡇࢀࡽࡢ࣮ࣜࢻࢆ Bismark ࡼ ࡗ࡚ mPing 㓄ิ࠾ࡼࡧࢿࢤࣀ࣒࣐ࢵࣆࣥࢢࡋࡓ ࡇࢁࠊ46,180 ࣮ࣜࢻ (0.03%)ࡀ mPing 㓄ิ࣐ࢵ ࣆࣥࢢࡉࢀࡓࠋࡇࢀࡽࡢ࠺ࡕࠊ5,652 ࣮ࣜࢻ (0.004%) ࡀ mPing ᤄධ⨨ࡢỴᐃ⏝࠸ࡿࡇࡀ࡛ࡁࡓࠋ mPing ࡣࠊရ✀㖟ᆓ࠾࠸࡚ࠊ1,000 ࢥࣆ࣮௨ୖ Ꮡᅾࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋᮏ◊✲࠾࠸࡚ࠊ106 ⟠ᡤࡢ mPing ᤄධࢆྠᐃࡋࠊࡇࢀࡽࡢ࠺ࡕ 21 ⟠ᡤ ࡢ mPing ࡢ࣓ࢳࣝ⛬ᗘࢆゎᯒ࡛ࡁࡓࠋࡑࡢ⤖ᯝࠊ mPing ෆ㒊ࡢࢩࢺࢩࣥࡢᖹᆒ࣓ࢳࣝ⛬ᗘࡣࠊCGࠊ CHG ࠾ࡼࡧ CHH ࢧࢺ࠾࠸࡚ࡑࢀࡒࢀ 95%ࠊ - 433 - බເᨭㄢ㢟㸸2010 ᖺᗘ ࣃ࣮ࢯࢼࣝࢤࣀ࣒ሗࡶ࡙ࡃ⬻ᝈࡢⓎᶵᵓࡢゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ ㎷ ┬ḟ࣭ᮾிᏛ་Ꮫ㒊㝃ᒓ㝔 㸺⫼ᬒ࣭┠ⓗ㸼 ձ㸬➽ⴎ⦰ᛶഃ⣴◳/ࣃ࣮࢟ࣥࢯࣥㄆ▱」ྜ ➽ⴎ⦰ᛶഃ⣴◳/ࣃ࣮࢟ࣥࢯࣥㄆ▱」ྜ 㸦ALS/PDC㸧ࡣࠊ㐠ືࢽ࣮ࣗࣟࣥ≧ຍ࠼ࣃ࣮࢟ ࣥࢯࢽࢬ࣒ࠊㄆ▱ࢆక࠸ࠊ⌮Ꮫⓗᗈ⠊࡞ tau ࡢỿ╔ࢆㄆࡵࡿ≉ᚩⓗ࡞ᝈ࡛࠶ࡿࠋࢢ࣒⣖ఀ ༙ᓥ㞟✚ᆅࡀ࠶ࡾࠊ⣖ఀ༙ᓥ࠾ࡅࡿᮏᝈࡣ⣖ ఀ ALS/PDC ࡤࢀ࡚࠸ࡿࠋᙉ࠸ᐙ᪘㞟✚ᛶࡀㄆ ࡵࡽࢀࠊ㑇ఏᏛⓗ⣲ᅉࡀⓎ㛵ࢃࡿ᥎ᐃࡉࢀ࡚ ࠸ࡿࠋᚑ᮶ࡽ㐃㙐ゎᯒࡀヨࡳࡽࢀ࡚࠸ࡿࡀࠊࣃࣛ ࣓ࢺࣜࢵࢡ㐃㙐ゎᯒࡣᅔ㞴࡛࠶ࡾࠊ㑇ఏᙧᘧࡀᚲࡎ ࡋࡶ᫂☜࡛࡞ࡃࠊ㧗㱋Ⓨ࡛࠶ࡾ phenocopy ࡢᏑᅾ ࢆࡣ㝖እ࡛ࡁࡎࠊᖖ㆑ⓗ࡞༢୍㑇ఏᏊᝈ ࡣ㝈ࡽ࡞࠸ࡇ➼ࡀ⌮⏤ࡋ࡚⪃࠼ࡽࢀ࡚ࡁࡓࠋ ᅇࠊᐙ⣔ᒓࡍࡿ 2 ྡࡢ๗᳨ࡘ࠸࡚ࠊࢤࣀ ࣒㓄ิゎᯒࢆ⾜ࡗ࡚࠸ࡓࡔࡁࠊALS/PDC ࡢཎᅉࡘ ࠸᳨࡚ウࢆ⾜ࡗࡓࠋ ղ㸬ከ⣔⤫ⴎ⦰ ከ⣔⤫ⴎ⦰ࡣ㸪⮬ᚊ⚄⤒㞀ᐖຍ࠼㸪ᑠ⬻ኻㄪ㸪 ࣃ࣮࢟ࣥࢯࣥ≧㸪㗽య㊰㞀ᐖ࡞ࡢ≧ࢆࡁࡓࡍ 㐍⾜ᛶࡢ⚄⤒ኚᛶᝈ࡛࠶ࡿ㸬ཎ๎ࡋ࡚ᏙⓎᛶ࡛ ࠶ࡾ㸪㑇ఏᛶࡢ࡞࠸ᝈ⌮ゎࡉࢀ࡚ࡁࡓࡀ㸪㏆ᖺ㸪 ከⓎᐙ⣔ࡀሗ࿌ࡉࢀࡿࡼ࠺࡞ࡾ㸪ᝈ㛵㐃㑇ఏᏊ ࡢ᥈⣴ࡀᗈࡃ⾜ࢃࢀࡿࡼ࠺࡞ࡗ࡚ࡁࡓ㸬ᚑ᮶㸪Ꮩ Ⓨᛶᝈࡢᝈឤཷᛶ㑇ఏᏊ᥈⣴ࡣ㸪㢖ᗘࡢ㧗࠸ ከᆺࢆ࣐࣮࣮࢝ᝈ⪅⩌࣭ᑐ↷⩌࡛ࣞࣝ㢖ᗘࡢ㐪 ࠸ࢆ᥈⣴ࡍࡿᡭἲ㸦ࢤࣀ࣒࣡ࢻ㛵㐃ゎᯒ㸧ࡀᗈࡃ ⾜ࢃࢀ࡚ࡁࡓ㸬ࡇࡢࡼ࠺࡞᪉ἲ᳨࡛ฟࡉࢀࡿᝈ㛵 㐃ከᆺࡢ࢜ࢵࢬẚࡣ 1㹼2 ಸ⛬ᗘ࡛࠶ࡿ㸬ࡑࡢ୍᪉㸪 ࣞࣝ㢖ᗘࡀ 1㸣⛬ᗘ㸪࠶ࡿ࠸ࡣࡉࡽ㢖ᗘࡢప࠸ rare variants ࡀᝈ㛵㐃ࡍࡿሙྜ㸪୍⯡࢜ࢵࢬ ẚࡀ 2㹼10 ಸᙉ࠸ᙳ㡪ᗘࢆᣢࡘᅉᏊࡀྠᐃࡉࢀࡿ ࡇࡀከ࠸㸬ࡇࡢࡼ࠺࡞ᅉᏊࡣ㞟ᅋෆࡢ㢖ᗘࡋ࡚ ࡣ⛥ࡔࡀ㸪ែᙉࡃᙳ㡪ࡍࡿᅉᏊ࡛࠶ࡾ㸪ែࡢ ゎ᫂ࡸ⒪ἲࡢ㛤Ⓨ࠸࠺Ⅼ࡛ᐤࡍࡿࡶࡢࡀࡁ ࠸⪃࠼ࡽࢀࡿ㸬ࡇࡢࡼ࠺࡞ rare variants ࢆ୰ᚰ ᝈ㛵㐃ኚ␗ࢆ᥈⣴ࡍࡿࡇࢆ┠ᶆ㸪ᅇ㸪 MSA ᝈ⪅ 14 㸪ᖖᑐ㇟⪅ 7 ࢆᑐ㇟࢚ࢡࢯ࣮࣒ゎᯒ ࢆ⾜ࡗ࡚࠸ࡓࡔࡁ᳨ウࡋࡓ㸬 㸺ᨭࡢෆᐜ㸼 ᨭࡼࡗ࡚㸪ձ㸬➽ⴎ⦰ᛶഃ⣴◳/ࣃ࣮࢟ࣥ ࢯࣥㄆ▱」ྜᝈ⪅ 2 ᳨యࡢࢤࣀ࣒ゎᯒ㸪࡞ࡽࡧ ղ㸬ከ⣔⤫ⴎ⦰ᝈ⪅ 21 ᳨యࡢ࢚ࢡࢯ࣮࣒ゎᯒࢆ ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓ㸬 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ձ㸬➽ⴎ⦰ᛶഃ⣴◳/ࣃ࣮࢟ࣥࢯࣥㄆ▱」ྜ ࢤࣀ࣒㓄ิゎᯒࡣ HiSeq2000 ࢆ⏝࠸࡚⾜ࢃࢀࠊ BWA ࡛ཧ↷㓄ิ hg19 alignment ࡉࢀࡓࠋྜィ࡛ 62.8Xࠊ72.9X ࡢ࢝ࣂࣞࢵࢪ࡞ࡾࠊ༑ศ࡞ࢹ࣮ࢱࡀ ᚓࡽࢀࡓ⪃࠼ࡽࢀࡓࠋSAMtools ࢆ⏝࠸ࡿࡇ࡛ࠊ ࡑࢀࡒࢀ 3604450ࠊ3576323 ಶࡢ୍ሷᇶ⨨ 587322ࠊ585433 ಶࡢᑠḞኻ࣭ᤄධࡀㄆࡵࡽࢀࡓࠋ ࠺ࡕࠊ㑇ఏᏊෆࡢ㠀ྠ⩏⨨ࢫࣉࣛࢫ㒊Ꮡ ᅾࡍࡿࡶࡢ࡛ࠊdbSNP Ꮡᅾࡋ࡞࠸ࡶࡢ⤠ࡿࠊ ୍ሷᇶ⨨ࡀ 347ࠊ326 ಶࠊᑠḞኻ࣭ᤄධࡀ 52ࠊ46 ಶㄆࡵࡽࢀࡓࠋࡑࡢᚋྠᵝ⾜ࢃࢀࡓࠊ᫂ࡽ࡞⾑ ⦕㛵ಀࡢ࡞࠸ 5 ๗᳨ࡢࢤࣀ࣒㓄ิゎᯒ࠶ࢃࡏ ࡚ゎᯒࡀ⾜ࢃࢀࡓࡀࠊ࡛ඹ㏻ࡍࡿ⩻ヂ㡿ᇦࡢ ኚ␗ࡣぢ࠸ࡔࡍࡇࡀ࡛ࡁ࡞ࡗࡓࠋᅇ๗᳨ ࠸࠺デ᩿ࡀ☜ᐃࡋࡓ㝈ࡗ࡚ゎᯒࢆ⾜ࡗࡓࡀࠊ ඹ㏻ࡍࡿኚ␗ࡀぢฟࡏ࡞ࡗࡓࡇࡽࠊ᳨ฟࡀẚ ㍑ⓗᅔ㞴࡛࠶ࡿᵓ㐀ኚ␗ࠊࣜࣆ࣮ࢺఙ㛗ࠊࣥࢺࣟ ࣭ࣥ㑇ఏᏊ㛫ࡢኚ␗࡞ࡢྍ⬟ᛶࢆ⪃࠼ࠊࡉࡽ᳨ ウࢆ⾜࠺ᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡓࠋ ղ㸬ከ⣔⤫ⴎ⦰ MSA ᝈ⪅ 14 㸪ᖖᑐ㇟⪅ 7 ࢆᑐ㇟࢚ࢡࢯ ࣮࣒ゎᯒࢆ⾜ࡗࡓ㸬ࡲࡎ㸪ᝈ㛵㐃ࡍࡿᙳ㡪ᗘࡢ ᙉ࠸ኚ␗ࡣ㸪㑇ఏᏊ⏘≀ࡢᶵ⬟ࢆኚࡉࡏࡿ㠀ྠ⩏ ኚ␗࡛࠶ࡿ࠸࠺௬ᐃࡢࡶ㸪㠀ྠ⩏ኚ␗ࢆᢳฟࡋ ࡓ㸬㠀ྠ⩏ኚ␗ࡣ㸪1 ࠶ࡓࡾ 10,000㹼12,000 ಶ⛬ ᗘྠᐃࡉࢀࡓ㸬21 㸪42 ࡛ࣞࣝ㞟ィࡋࡓࡇࢁ㠀 ྠ⩏ኚ␗ࡣ 29,359 ✀㢮ࡔࡗࡓ㸬42 ࣞࣝ୰ 1 ࣞ ࣝࡢ㢖ᗘࡢኚ␗ࡣ 13,345 ✀㢮㸦45.5㸣㸧㸪ࡲࡓࣞ ࣝ㢖ᗘ 1㸣ᮍ‶ࡢኚ␗ࡣ 18,850 ✀㢮㸦64.2%㸧㐣 ༙ᩘࢆ༨ࡵࡿࡇࡀศࡗࡓ㸬ࣞࣝ㢖ᗘ 1㸣ᮍ‶ࡢ ኚ␗ࢆᑐ㇟㸪࣑ࣀ㓟ኚࡢᶵ⬟ኚࡢண ࣝ ࢦࣜࢬ࣒ࢆ⏝࠸࡚㸪ᝈ㛵㐃ࡍࡿኚ␗ࡢೃ⿵ࢆ⤠ ࡿࡇࢆヨࡳࡓ㸬dbNSFPver1.1 ࢆ⏝࠸࡚㸪 Polyphen2, SIFT, MutationTaster, LRT, Phylop ࡑ ࢀࡒࢀࡢண ࢆࢫࢥࡋ㸪ࢫࢥࡢ㧗࠸ኚ␗㸦5 Ⅼ‶Ⅼ୰ 4 Ⅼ௨ୖ㸧ࢆᢳฟࡋࡓࡇࢁ㸪18,850 ✀㢮 ୰ 4,028 ✀㢮ࡢኚ␗⤠ࡽࢀࡓ㸬ᅇࡢᨭࡢ⠊ᅖ ࡛࠶ࡿ MSA ᝈ⪅ 14 㸪 ᖖᑐ㇟⪅ 7 ࡢẚ㍑࡛ࡣ㸪 ᭷ព㛵㐃ࡍࡿኚ␗ࡣㄆࡵࡽࢀ࡞ࡗࡓࡀ㸪ᩘࢆ ቑࡸࡋ࡚ᘬࡁ⥆ࡁ᳨ウࢆ⥆ࡅ࡚࠸ࡃ㸬 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ձ㸬➽ⴎ⦰ᛶഃ⣴◳/ࣃ࣮࢟ࣥࢯࣥㄆ▱」ྜ ୍⯡࣓ࣥࢹࣝ㑇ఏᙧᘧࡀ௬ᐃ࡛ࡁࡿᐙ⣔ࡢᐙ ⣔ゎᯒ࠾࠸࡚ࡣ㸪ࣃ࣓ࣛࢺࣜࢵࢡ㐃㙐ゎᯒࡼࡗ ࡚ೃ⿵㡿ᇦࢆ⤠ࡾࡇࡴࡇࡀ࡛ࡁࡿ㸬ࡋࡋ㸪ᅇ ᨭ㡬࠸ࡓ ALS/PDC ࡢࡼ࠺࡞㸪㑇ఏᙧᘧࡀᚲࡎࡋ ࡶ᫂☜࡛࡞ࡗࡓࡾ㸪㧗㱋Ⓨ࡛ phenocopy ࡢᏑᅾ ࢆࡣ㝖እ࡛ࡁ࡞ࡗࡓࡾࡍࡿࡼ࠺࡞ᐙ⣔ࡢゎ ᯒ࡛ࡣ㸪ᵝࠎ࡞ྍ⬟ᛶࢆ௬ᐃࡋ࡚ࢤࣀ࣒࣡ࢻ᳨ ウࡍࡿࡇࡀᚲせ࡛࠶ࡿ㸬ᅇࡢ᳨ウ࡛ࡣ㸪⩻ヂ㡿 ᇦෆࡢሷᇶ⨨ኚ␗㸪short indel ࡋ᳨࡚ฟࡉࢀࡓ ኚ␗ࡢ⠊ᅖ࡛ࡣ㸪ೃ⿵ࡋ࡚ṧࡿኚ␗ࡣぢฟࡏ࡞ ࡗࡓࡓࡵ㸪ࡑࡢࡢྍ⬟ᛶ㸦ᵓ㐀ኚ␗㸪ࣜࣆ࣮ࢺఙ 㛗㸪ࣥࢺ࣭ࣟࣥ㑇ఏᏊ㛫ࡢኚ␗㸧ࡘ࠸᳨࡚ウࡍ - 434 - ࡿᚲせࡀ♧၀ࡉࢀࡓ㸬ALS/PDC ࡢࡼ࠺ᚑ᮶ࡢ᪉ἲ ࡛ࡣࣉ࣮ࣟࢳࡀᅔ㞴࡞ᐙ⣔ᑐࡋ࡚㸪⥙⨶ⓗ࡞ ࢤࣀ࣒ゎᯒࢆ⾜࠺ࡇ࡛ึࡵ࡚ᅉゎ᫂ࡢᒎᮃࡀᣅ ࡅࡓゝ࠼ࡿ㸬 ղ㸬ከ⣔⤫ⴎ⦰ ᏙⓎᛶᝈࡘ࠸࡚ࡣ㸪ᚑ᮶㸪㢖ᗘࡢ㧗࠸ከᆺࢆ ࣐࣮࣮࢝ᝈ⪅⩌࣭ᑐ↷⩌࡛ࣞࣝ㢖ᗘࡢ㐪࠸ࢆ᥈ ⣴ࡍࡿᡭἲ㸦ࢤࣀ࣒࣡ࢻ㛵㐃ゎᯒ㸧ࡀᗈࡃ⾜ࢃࢀ ࡚ࡁࡓ㸬ࡇࡢ᪉ἲࡣ㸪㞟ᅋෆࡢ㢖ᗘࡀ㧗ࡃ㸪ᙳ㡪ᗘ ࡢẚ㍑ⓗᙅ࠸ኚ␗ࢆᗈࡃ᳨ฟࡍࡿࡇࡀ࡛ࡁࡿࡀ㸪 㢖ᗘࡀపࡃ㸪ᙳ㡪ᗘࡢᙉ࠸㑇ఏᅉᏊࡣ᳨ฟࡀᅔ㞴ࡔ ࡗࡓ㸬࢚ࢡࢯ࣮࣒ゎᯒࢆ⾜࠺ࡇ࡛ึࡵ࡚㸪㢖ᗘࡢ ప࠸ኚ␗ࢆྵࡴ⥙⨶ⓗ࡞⩻ヂ㡿ᇦෆࡢኚ␗ྠᐃࡀྍ ⬟࡞ࡾ㸪ᏙⓎᛶᝈࡢ㑇ఏᅉᏊᐜゎ᫂ࡘ࡞ࡀ ࡿᒎᮃࡀᣅࡅࡓ㸬ᅇࡢᨭࡢ⠊ᅖ࡛ࡣᩘࡀᑡ࡞ ࡃ㸪༢⊂࡛⤖ㄽࢆᚓࡿࡇࡣ㞴ࡋ࠸ࡀ㸪ᚋࡢ㛵㐃 ゎᯒࡢࢹࢨࣥᚲせ࡞ࢹ࣮ࢱࡀᚓࡽࢀࡓⅬࡣࡁࢃ ࡵ࡚᭷⏝ࡔࡗࡓ㸬 ࡞࠾㸪ᨭࡢᑐ㇟ࡢ┤᥋ⓗ࡞ᡂᯝ࡛ࡣ࡞࠸ࡀ㸪ᨭ ࡢࢹ࣮ࢱࢆཧ⪃ࡋ࡚㸪◊✲ࢆⓎᒎࡉࡏࡓ⤖ᯝ㸪 ᐙ⣔࡞ࡽᚓࡽࢀࡓೃ⿵㑇ఏᏊࣉ࣮ࣟࢳࡼ ࡗ࡚㸪㢖ᗘࡀపࡃ㸪ᙳ㡪ᗘࡢᙉ࠸ከ⣔⤫ⴎ⦰ࡢ㑇 ఏᅉᏊࡋ࡚㸪COQ2 㑇ఏᏊ㸦2013 ᖺ㸧㸪GBA 㑇ఏ Ꮚ㸦2015 ᖺ㸧ࢆྠᐃࡋሗ࿌ࡋ࡚࠸ࡿ㸬 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࣭The Multiple-System Atrophy Research Collaboration (Mitsui J et al.,) N Engl J Med. 2013; 369: 233-44. ࣭Mitsui J, Tsuji S. Biochem Biophys Res Commun. 2014; 452: 221-5. ࣭Mitsui J et al., Ann Clin Transl Neurol. 2015; 2: 417-426 - 435 - බເᨭㄢ㢟㸸2010 ᖺᗘ ⎔ቃởᰁ≀㉁ῧຍᅵተ࡛ࡢᚤ⏕≀㞟ᅋᛂ⟅ᵝᘧࡢ◊✲ ᇶ┙◊✲㸦㹀㸧 ὠ⏣ 㞞Ꮥ࣭ᮾᏛᏛ㝔⏕⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ᭷ᐖᏛ≀㉁࡛ởᰁࡉࢀࡓᅵተࡢίࡣᙜヱᅵ ተ᳇ᜥ⣽⳦㞟ᅋࡀࡁࡃᐤࡍࡿࠋᮏ⏕ែ⣔࡛ࡢở ᰁ≀㉁ศゎ㛵ࡍࡿ⣽⳦⩌ࡸࡑࡢởᰁ≀㉁ศゎ㓝 ⣲⩌ࡢᥦ♧ࢆࡵࡊࡋࠊ4 ✀⎔ቃởᰁ≀㉁[3-ࢡࣟࣟᏳ ᜥ㤶㓟(3CB)ࣇ࢙ࢼࣥࢺࣞࣥ(Phn)ࢆྵࡴ 3 ✀ከ⎔ ⰾ㤶᪘ྜ≀(PAHs)]ࢆྠῧຍࡋࡓ㛢㙐⣔ᅵተ(௨ ୗࠕඛ⾜ởᰁᅵተࠖ⾲グ)ࢆ⏝࠸࡚ࠊ⣽⳦㞟ᅋࡢ⳦ ྀ㑇ఏᏊࣉ࣮ࣝࡢ⤒ⓗኚືᵝᘧࡢ࣓ࢱࢤࣀ࣒ゎ ᯒྛ✀⳦ᰴ࣭ศゎ㓝⣲㑇ఏᏊ༢㞳࣭ゎᯒࢆᐇࡋ ࡚ࡁࡓࠋᮏ◊✲࡛ࡣࠊୖグᅵተ࡛ 3CB Phn ࡢศゎ ୰ᚰⓗᙺࢆᯝࡓࡋ࡚࠸ࡓྛࠎࡢ⣽⳦ᒓゎ᫂ࢆࡵ ࡊࡋࡓࠋࡉࡽࠊ㠀ởᰁྠ୍ᅵተࡢ⁛⳦ヨᩱඛ ⾜ởᰁᅵተ⏤᮶⣽⳦㞟ᅋࢆࠕ⛣᳜ࠖࡋࠊ3CB ༢⊂ῧ ຍᚋࡢ⳦ྀ࣭㑇ఏᏊࣉ࣮ࣝࡢ⤒ⓗኚືࢆ᳨ウࡋࠊ 3CB ศゎ⣽⳦ᒓࡢᙺ᳨ドࢆࡵࡊࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ඛ⾜ởᰁᅵተ⏤᮶࡛ࠊởᰁᚋึᮇ୍ⓗඃ༨ ࡋࡓ Burkholderia ᒓ 6 ᰴࠊࡑࡋ࡚ࠊPhn ศゎ⬟ࢆ ♧ࡋࡓ Mycobacterium ᒓᰴ Paenibacillus ᒓᰴࡢࢻࣛ ࣇࢺࢤࣀ࣒㓄ิỴᐃ(Roche 454)ࠋ(2)ୖグ⁛⳦ᅵተ ⛣᳜ࡋࡓ⣽⳦㞟ᅋ 3CB ῧຍᚋࡢ」ᩘⅬ⏤᮶࣓ ࢱࢤࣀ࣒ࢆ⏝࠸࡚ࠊ⣽⳦㞟ᅋࡢ⳦ྀᥦ♧⏝ࡢ 16S rRNA 㑇ఏᏊ㓄ิỴᐃ(Roche 454)㑇ఏᏊࣉ࣮ࣝᥦ ♧⏝ࡢࢩࣙࢵࢺ࢞ࣥ㓄ิỴᐃ(Illumina HiSeq 2000)ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ởᰁᚋ 1 㐌㛫࡛ 3CB ࡀᾘኻࡋࠊ 3 ✀ PAHs ࡀ 3 㐌┠௨㝆ศゎ㛤ጞࡋࡓඛ⾜ởᰁᅵተࡢ࣓ࢱࢤࣀ ࣒ゎᯒ࡛ࠊE-Proteobacteria ⥙ࡢ Burkholderia ᒓ⣽⳦ ࡢ 1 㐌┠࡛ࡢᛴ⃭࡞ቑຍࡀぢฟࡉࢀࠊ3CB ศゎࡢ E-Proteobacteria ࡢ㛵 Phn ከẁ㝵ศゎࡢ๓༙ࡢ E-Proteobacteria ⥙ᚋ༙ࡢ Actinobacteria 㛛ࡢ⣽⳦ ࡢ㛵ࡀ♧၀ࡉࢀ࡚࠸ࡓࠋ ࠕᨭ࡛ࠖᚓࡓ Burkholderia ᒓ⣽⳦ 3 ᰴ Mycobacterium sp. EPa45 ᰴࡢࢻࣛࣇࢺ ࢤࣀ࣒ࢆࣞࣇࣞࣥࢫࢤࣀ࣒ࡋࡓඛ⾜ởᰁᅵተࡢ ࣓ࢱࢤࣀ࣒ゎᯒ࡛ྛởᰁ≀㉁ศゎ㛵ࢃࡿ⣽⳦ࢆ ᒓ࡛ࣞ࣋ࣝᥦ♧ྍ⬟࡞ࡾࠊ௨ୗࡢᡂᯝࢆᚓࡓࠋ(1-1) 3CB ศゎࡢ Burkholderia ᒓ⣽⳦ࡢ㛵ࠊࡉࡽ Phn ศ ゎ ࡢ ๓ ༙ ᚋ ༙ ࡢ Mycobacterium ᒓ Burkholderia ᒓࡢྛ⣽⳦ࡢ㛵ࠊࡀᙉࡃ᥎ᐃࡉࢀࡓࠋ (1-2)ࢻࣛࣇࢺ㓄ิࢆඖ Mycobacterium EPa45 ᰴࢤ ࣀ࣒㓄ิࢆỴᐃࡋࠊPhn ศゎ⤒㊰ศゎ㓝⣲ 㑇ఏᏊ⩌ࢆ♧ࡋࡓࠋࡉࡽᙜヱ㑇ఏᏊ⩌ᴟࡵ࡚┦ ྠᛶࡢ㧗࠸࣓ࢱࢤࣀ࣒㓄ิࡀඛ⾜ởᰁᅵተ࡛ Phn ศ ゎ ㇏ ᐩ Ꮡ ᅾ ࡋ ࠊ Phn ศ ゎ ๓ ༙ ẁ 㝵 ࡢ Mycobacterium 㛵ࢆ᥎ᐃ࡛ࡁࡓࠋ(1-3)ඛ⾜ởᰁᅵተ ࣓ࢱࢤࣀ࣒ࡽᇵ㣴㠀౫Ꮡⓗᡭἲ࡛」ᩘྲྀᚓࡋࡓⰾ 㤶᪘ྜ≀ศゎ㓝⣲㑇ఏᏊ⩌ࡢ୍㒊ࡣ Burkholderia ⏤᮶࡛ởᰁᅵተࡢࡳ࡛≉␗ⓗቑຍࡋ࡚࠸ࡓࠋ 㠀ởᰁྠ୍ᅵተࡢ⁛⳦ヨᩱඛ⾜ởᰁᅵተ⏤᮶ ⣽⳦㞟ᅋࢆ⛣᳜ࡋࡓ┤ᚋࡣつᶍ࡞⳦ྀኚືࡀ㉳ ࡁࡓࡀࠊኚື㙠㟼ࡢ 4 㐌┠ῧຍࡋࡓ 3CB ࡣ 1 㐌 㛫࡛ᾘኻࡋࡓࠋ⛣᳜⏝ࡋࡓ⣽⳦㞟ᅋࠊ⛣᳜ᚋࡢ 2 Ⅼࡢ⣽⳦㞟ᅋࠊࡑࡋ࡚ࠊ3CB ởᰁᚋࡢ 3 Ⅼࡢ⣽ ⳦㞟ᅋࡽㄪ〇ࡋࡓ࣓ࢱࢤࣀ࣒ࢆ⏝࠸࡚ࠊ⳦ྀ㑇 ఏᏊࣉ࣮ࣝࡢኚືࢆ᳨ウࡋࠊ(2-1)⛣᳜┤ᚋࡢ㞟ᅋ࡛ ࡢ Burkholderia ᒓࢆྵࡴ୍㒊ᒓ⩌ࡢ༶ᗙඃ༨ࠊ ࡑࡢᚋࡢ⛣᳜࣐ࢼ࣮ᒓቑຍࢆకࡗࡓ⳦ྀ⤌ᡂࡢ Ᏻᐃഴྥ(ࡓࡔ⛣᳜ᚋ 4 㐌Ⅼ࡛ࡶࠊࡲࡔ⳦ྀኚື ࠶ࡾ)ࢆ♧ࡍࡇࠊ(2-2) 3CB ༢⊂ởᰁࡼࡾࠊ㞟ᅋෆ Burkholderia ᒓ⣽⳦ࡣඛ⾜ởᰁᅵተࡢึᮇ㐣⛬ྠ ᵝࡢ⤒ⓗኚືᵝᘧࢆ♧ࡋࠊBurkholderia ᒓ⏤᮶ࡢ 3CB ศゎ㓝⣲㑇ఏᏊ㔞ࡀቑࡋࡓࠋ௨ୖࡼࡾࠊඛ⾜ ởᰁᅵተ࡛ࡢ Burkholderia ᒓ⣽⳦ࡢ 3CB ศゎࡢ㔜 せᛶࢆ᫂♧࡛ࡁࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ୖ㏙ࡢࡼ࠺ඛ⾜ởᰁᅵተ࡛ࡢຠ⋡ⓗ Phn ศ ゎࡣ Mycobacterium ᒓ⣽⳦ Burkholderia ᒓ⣽⳦ࡢ ࠕ༠ㄪ௦ㅰࠖࡀᙉࡃ᥎ᐃࡉࢀࡓࡀࠊởᰁᚋ༙ᖺ㛫࡛ Mycobacterium ᒓ⣽⳦ࡣ㞟ᅋࡢ 1㸣௨ୗ࡛࠶ࡗࡓࡓࡵࠊ Burkholderia ᒓ⣽⳦ࡣ Mycobacterium ᒓ⣽⳦ࡢ Phn ศゎ⬟ࢆ༠ㄪ௦ㅰࡣูᶵᵓ࡛ᙉࡉࡏࡓྍ⬟ᛶ ࡀ♧၀ࡉࢀࡓࠋ୍᪉ࠊᙜヱᅵተࡽࡢ Phn ศゎ Mycobacterium EPa45 ᰴ༢㞳㐣⛬࡛ࠊᮏᰴᅵተ᳇ᜥ Burkholderia ᒓ⣽⳦ࡢᙉ࠸ඹᏑ㛵ಀࢆぢฟࡋࠊ๓⪅ ᰴࡀ♧ࡍ Phn ẘᛶࢆඹᏑ Burkholderia ᒓ⣽⳦ᰴࡀ⦆ ࡍࡿຠᯝࢆぢ࠸ࡔࡋࡓࠋᮏຠᯝࡢศᏊᶵᵓゎ᫂ࢆ ⌧ᅾ᳨ウࡋ࡚࠸ࡿࡀࠊศゎ⳦ඹᏑࡍࡿ㠀ศゎ⳦ࡢ ᙺࡑࡢᶵᵓࡀゎ࡛᫂ࡁࠊ⮬↛⏕ែ⣔࡛ࡢ⎔ቃở ᰁ≀㉁ศゎᵝᘧࡢᐇయࡢᥦ♧⧅ࡆࡓ࠸ࠋ⁛⳦ᅵ ተࡢ⣽⳦㞟ᅋ⛣᳜ᚋࡢ⤒ⓗ࣓ࢱࢤࣀ࣒ゎᯒࢆ ࠕᨭࠖ౫㢗ࡋࡓࡀࠊྠ୍⁛⳦ᅵተࢆศࡋࡓ 」ᩘヨᩱ࡛ࡢ⊂❧᳨ウࢆᅇ㑊ࡋࡓࡓࡵࠊ」ᩘヨᩱ㛫 ࡛ࡢ⳦ྀኚືࡢ⌧ᛶࡣ᫂ࡔࡗࡓࠋࡑࡢᚋࡢᡃࠎ ࡢ」ᩘヨᩱ࡛ࡢ⳦ྀኚືࡢ㛗ᮇ㛫᳨ウ࡛ࠊ⛣᳜ᚋ༙ ᖺ࡛Ᏻᐃ⮳ࡿࡲ࡛ࡢ⳦ྀࡢኚືᵝᘧ⤌ᡂࡣヨ ᩱ㛫࡛ᕪ␗ࡀ࡞ࡗࡓࠋᮏ▱ぢࡣࠊᚋணᐃࡋ࡚࠸ ࡿ㛢㙐ᅵተ⣔⣽⳦㞟ᅋࡢ⳦ྀ࣭㑇ఏᏊࣉ࣮ࣝኚືࢆ つᐃࡍࡿ⎔ቃせᅉࡢ᫂♧ᙜヱኚືᶵᵓࡢゎ᫂㛵 ࡋ࡚ࠊ⌧ᛶࡢ㧗࠸ᡂᯝࢆᚓࡿᇶ┙࡞ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ຍ⸨ࡽ, J. Environ. Biotechnol. 10: 63-71 (2010) ỌᒣࡽJ. Environ. Biotechnol. 13: 51-56 (2013) Mori, H. et al., DNA Res. 21: 217-227 (2014) Nagayama, H. et al., Appl. Microbiol. Biotechnol. 99: 4453-4470 (2015) Kato, H. et al., Genome Announc. 3: e00782-15 (2015) ຍ⸨ࡽ. ᪥ᮏᚤ⏕≀⏕ែᏛㄅ 30: 57-64 (2015) Kato, H. et al., DNA Res. 22: 413-424 (2015) - 436 - බເᨭㄢ㢟㸸2011~2012 ᖺᗘ 㐲㝸ࢤࣀ࣒㒊㛫ࡢ┦స⏝࠾ࡅࡿ㹇㹇ᆺ㹂㹌㸿ࢺ࣏ࢯ࣓࣮ࣛࢮࡢᙺ ᇶ┙◊✲㸦㹀㸧 ⟄ ◊࣭ᒸᒣᏛ⮬↛⏕⛉Ꮫ◊✲ᨭࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ௨๓ᡃࠎࡣ '1$ ࢺ࣏ࢯ࣓࣮ࣛࢮ ,,ș㸦ࢺ࣏ ,,ș㸧 ࡀ⚄⤒⣽⬊ࡢ⤊ᮎศ࠾࠸୍࡚⩌ࡢ⚄⤒㛵㐃㑇ఏ Ꮚࡢ㌿ࢆㄏᑟࡋࠊᡂ⇍ࢽ࣮ࣗࣟࣥࡢᡂᚲ㡲 ࡞ࡗ࡚࠸ࡿࡇࢆ♧ࡋࡓࠋᮏ◊✲࡛ࡣࠊࢺ࣏ ,,șࡢ స⏝Ⅼࢆࢤࣀ࣒ '1$ ୖ࣐ࢵࣆࣥࢢࡍࡿ᪉ἲࢆ㛤Ⓨ ࡋࠊࡇࡢ㓝⣲ࡀᇶ㉁ࡍࡿ ᮏࡢ '1$ 㙐ࡀ㐲㝸ࡢࢤ ࣀ࣒㒊⏤᮶ࡍࡿࢣ࣮ࢫࡢᏑᅾࢆド᫂ࡍࡿࡶ ࠊࡑࡢ⤖ᯝ⏕ࡌࡿࢡ࣐ࣟࢳࣥ㧗ḟᵓ㐀ࡢኚࡀ㌿ ࢆไᚚࡍࡿ࠸࠺௬ㄝࢆ᳨ドࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ⿕ᨭ⪅ࡢ◊✲ᐊ࡛ㄪ〇ࡋࡓ⣽⬊ '1$ ᩿∦ࢆ㏦ࡋࠊ ࡍ࡚᪂ୡ௦ࢩ࣮ࢡ࢚ࣥࢧ㸦+L6HT㸧ࡼࡾ࣌ ࢚ࣥࢻ࡛ㄞࢇࡔ࣮ࣜࢻ㓄ิࢆ㏉ࡋ࡚ࡶࡽࡗࡓࠋලయ ⓗࡣ௨ୗ㸲✀㢮ࡢࢧࣥࣉࣝࡢゎᯒࢆ౫㢗ࡋࡓࠋ㸯㸧 51$VHT ⏝ࡢ 51$㸦ィ㸱࣮ࣞࣥศ㸧㸰㸧ࢺ࣏ ,,șస⏝ Ⅼ㸦H7,3VHT㸧⏝ࡢ㸰ࡘࡢ '1$ ⏬ศ㸦ィ㸴࣮ࣞࣥศ㸧 㸱㸧㐲㝸స⏝㒊ゎᯒ㸦H7,33(6㸧⏝ࡢ࣓࢟ࣛ '1$ ᩿∦㸦ィ㸶࣮ࣞࣥศ㸧ࠋ㸲㸧ࢡ࣐ࣟࢳࣥ㧗ḟᵓ㐀ゎᯒ 㸦)$,5(VHT㸧⏝ࡢ '1$㸦ィ㸴࣮ࣞࣥศ㸧ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ,, ᆺࢺ࣏ࢯ࣓࣮ࣛࢮࡢヨ㦂⟶ෆᛂ࣓࢝ࢽࢬ࣒ ࡘ࠸࡚ࡣ᪤ࡼࡃ⌮ゎࡉࢀ࡚࠸ࡓࡀࠊࡇࡢ㓝⣲ࡀ⏕ ⣽⬊ࡢ᰾ෆ࡛㐲ࡃ㞳ࢀࡓࢤࣀ࣒㒊㛫ࡢ┦స⏝ࢆ ፹ࡍࡿྰࡘ࠸࡚ࡣࠊỌࡽࡃ᫂ࡔࡗࡓࠋᮏ ◊✲ࡢ┠ⓗࡣࠊࡇࡢၥ㢟ࢆゎỴࡍࡿࡓࡵᚲせ࡞ ᢏ⾡ H7,33(6 ࢆ㛤Ⓨࡋࠊ ࡇࢀࢆ᳨ドࡍࡿࡇ࡛࠶ࡿࠋ ሷᇶ㓄ิሗࢆぢ࡞ࡀࡽ᪂ࡓ࡞ᐇ㦂᮲௳ࢆヨࡳࡿ ࠸࠺ࠊヨ⾜㘒ㄗࡢ⧞ࡾ㏉ࡋ࡛࠶ࡗࡓࡀࠊ᭱⤊ⓗࡣ ᭱ᑠ㝈ࡢᐇ㦂ࡼࡗ࡚᭱㝈ࡢᡂᯝࢆᚓࡿࡇࡀ࡛ ࡁࡓࠋ௨ୗ࡞ᡂᯝࢆᣲࡆࡿࠋ㸯㸧P51$VHT ࡢࢹ ࣮ࢱࢆゎᯒࡋࠊศࡀ㐍ࡴࢺ࣏ ,,șࡢάᛶ౫Ꮡ ࡋ࡚Ⓨ⌧ࡀㄏᑟࡉࢀࡿ㑇ఏᏊ⩌㸦$ 㑇ఏᏊ㸧ᢚไ ࡉࢀࡿ㑇ఏᏊ⩌㸦& 㑇ఏᏊ㸧ࡢᏑᅾࢆ᫂ࡽࡋࡓࠋ *2 ゎᯒࡼࢀࡤࠊࡇࢀࡽࡢ㑇ఏᏊ⩌௨እࡶᶵ⬟ⓗ ≉ᚩࢆࡶࡘⓎ⌧ࢢ࣮ࣝࣉࡀྠᐃࡉࢀࡓࠋⓎ⌧ࢢ࣮ࣝ ࣉࡈᶵ⬟ࢆㄪࡿࠊ$ 㑇ఏᏊࡣᡂ⇍⚄⤒⣽⬊ ࡛ࠊ& 㑇ఏᏊࡣึᮇࡢ⚄⤒ศ㐣⛬࡛ാࡃ㑇ఏᏊࡀ ᭷ពከࡃࠊࢺ࣏ ,,șࡀ⤊ᮎศ࠾࠸୍࡚⩌ࡢ㑇 ఏᏊࡢⓎ⌧ࢆྜ⌮ⓗ୧᪉ྥไᚚࡋ࡚࠸ࡿࡇࡀ ศࡗࡓࠋࡇࢀࡽࡢ㑇ఏᏊࡀ༨ࡵࡿࢤࣀ࣒࣏ࢪࢩࣙ ࣥࡘ࠸࡚ࡶࠊ$ & ࡣࡑࢀࡒࢀ࡛㏆ࡃ㞟ࡲࡿࡀࠊ ࠸┦ᡭࢆ㝖ࡍࡿഴྥࡀ࠶ࡿ࠸࠺≉ᚩࡀㄆࡵ ࡽࢀࡓࠋࡲࡓࠊ$ 㑇ఏᏊࡣ㛗࠸㑇ఏᏊ㛗ࢆࡶࡘ㑇 ఏᏊࡀከࡃྵࡲࢀࠊࡇࢀࡽࡣ㛗࠸㑇ఏᏊ㛫㡿ᇦ㞄 ᥋ࡍࡿ࠸࠺ࠊ㢧ⴭ࡞≉ᚩࢆ♧ࡋࡓࠋ㸰㸧ࢺ࣏ ,,ș స⏝Ⅼ㸦ࢺ࣏ࢧࢺ㸧ࢆ⥙⨶ⓗゎᯒࡋ㸦H7,3VHT㸧 ࠊ 㸱ࡘࡢࢡࣛࢫ㸦7V7V7V㸧ศ㢮ࡋࡓࠋ7V ࡣ ㌿㛤ጞ㒊㸦766㸧㏆ഐ㑇ఏᏊᐩࡴࢤࣀ࣒㡿ᇦ ᗈࡃศᕸࡍࡿࠋ7V 7V ࡣ㑇ఏᏊஈࡋ࠸㑇ఏᏊ 㛫㡿ᇦከࡃࠊẚ㍑ⓗᑡᩘࡢᐦ㞟㒊㸦࣍ࢵࢺࢫ࣏ ࢵࢺ㸧ࢆᙧᡂࡋ࡚࠸ࡓࠋ㸱㸧H7,33(6 ࡽᚓࡽࢀࡿ ࢹ࣮ࢱࡣࠊ㸰⟠ᡤ⏤᮶ࡍࡿ '1$ ᩿∦ࡀࢲࣉࢱ࣮ ࢆࡋ࡚⤖ྜࡋࡓ࣓࢟ࣛ㓄ิࡽᡂࡿࠋ᭱ࡢၥ㢟 Ⅼ࡛࠶ࡗࡓࣀࢬࡢ㝖ཤࡣࠊ࣮ࣜࢻᩘࢆቑຍࡉࡏࠊ ୧➃ࡀࢺ࣏ࢧࢺ୍⮴ࡍࡿ࣓࢟ࣛࡢࢩࢢࢼࣝࢆ㑅 ᢥࡍࡿࡇࡼࡾゎỴࡉࢀࡓࠋ୍⮴ࡍࡿࢺ࣏ࢧࢺ ࡢ✀㢮ࡼࡗ࡚࣓࢟ࣛࢆ ✀㢮㸦7V7V7V ࣓࢟ ࣛ㸧ศ㢮ࡋࡓࠋ࣓࢟ࣛࡢゎᯒࡽᚓࡽࢀࡿࡍ࡚ ࡢሗ㸦㛗ࡉࠊ࣮ࣜࢻࡢ᪉ྥࠊࢤࣀ࣒࣏ࢪࢩࣙࣥ㸧 ࢆ⏝࠸ࠊ7V ࣓࢟ࣛࡣ㏆ࡃࡢ '1$ 㙐㛫ࡢᛂ㸦363㸧ࠊ 7V7V ࣓࢟ࣛࡣ㐲ࡃ㞳ࢀࡓ '1$ 㙐㛫ࡢᛂ㸦'63㸧 ࢆᫎࡋ࡚࠸ࡿࡇࢆ᫂ࡽࡋࡓࠋ7V7V ࢺ࣏ࢧ ࢺࡢ࣍ࢵࢺࢫ࣏ࢵࢺ୍⮴ࡋ࡚ࠊࡑࡢෆ㒊࠾ࡼࡧ ࣍ࢵࢺࢫ࣏ࢵࢺ㛫ከࡃࡢ࣓࢟ࣛࡀ࣐ࢵࣉࡉࢀࡿࡇ ࡽࠊࡇࢀࡽࡢ㒊ࡀ '63 ࣍ࢵࢺࢫ࣏ࢵࢺࢆᙧᡂ ࡋ࡚࠸ࡿࡇࡀุ᫂ࡋࡓࠋ㸲㸧'63 ࣍ࢵࢺࢫ࣏ࢵࢺࡢ ୍㒊ࡣࠊ┦ྠ㑇ఏᏊࣇ࣑࣮ࣜࡢࢡࣛࢫࢱ࣮ࡸࠊ ࢭࣥࢺ࣓ࣟࢆᵓᡂࡍࡿ㓄ิ㸦6DW,㸧ࡀ㧗㢖ᗘ Ꮡᅾࡋࡓࠋࢭࣥࢺ࣓ࣟࡀࢺ࣏ ,,ș ࡢࡓࡿస⏝ Ⅼࡢ୍ࡘ࡛࠶ࡿࡇࡣࠊ6DW,ࢆࣉ࣮ࣟࣈࡍࡿ㸱 '),6+㸭ࢺ࣏ ,,ș 㔜ᰁⰍࡼࡗ࡚☜ㄆࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 㸯㸧ᮏ◊✲ᨭࡼࡗ࡚ྍ⬟࡞ࡗࡓ᩿➃㓄ิࡢ つᶍࢩ࣮ࢡ࢚ࣥࢩࣥࢢࡣࠊ,, ᆺ '1$ ࢺ࣏ࢯ࣓࣮ࣛ ࢮ࡛ᐃࡉࢀࡿ㸰ࡘࡢᛂᵝᘧ㸦363 '63㸧ࡀ⏕⣽ ⬊ࡢ᰾ෆ࡛ᐇ㝿㉳ࡇࡗ࡚࠸ࡿࡇࢆド᫂ࡋࠊࢤࣀ ࣒ୖࡢ㒊ࢆࡶ≉ᐃࡋࡓࠋࡇࡢ㓝⣲ࡢ⏕⌮ᶵ⬟㛵 ࢃࡿ▱ぢࢆ᱁ẁ㐍ࡵࡿࣈࣞࢡࢫ࣮ࣝゝ࠼ࡿࠋ 㸰㸧ࢺ࣏ ,,ș ࡛ไᚚࡉࢀࡿ㑇ఏᏊࡋ࡚ࠊ⚄⤒ศ ࡢ␗࡞ࡿᮇⓎ⌧ࡍࡿ ⩌ࡢ⚄⤒㛵㐃㑇ఏᏊࡀᏑ ᅾࡋࠊ᭱⤊ศࢆቃࡋ࡚Ⓨ⌧ࡀᢚไࡉࢀࡿ㑇ఏᏊ ⩌ࡽㄏᑟࡉࢀࡿ㑇ఏᏊ⩌ࡢษࡾ᭰࠼ࢆࢺ࣏ ,,ș ࡀ⾜ࡗ࡚࠸ࡿࡇࡀ᫂☜࡞ࡗࡓࠋ⚄⤒ศ࠾ࡅ ࡿࡇࡢ㓝⣲ࡢ㔜せᛶࢆ᭦㏣ồࡍࡿ◊✲ࡀᮃࡲࢀࡿࠋ 㸱㸧ࢺ࣏ ,,ș ࡼࡿ㑇ఏᏊⓎ⌧ไᚚࡋ࡚ࠊ㸰ࡘࡢ ᶵᵓࡀ♧၀ࡉࢀࡓࠋ㑇ఏᏊࡢ㌿㡿ᇦ࡛㉳ࡇࡿ 363 ࡛ࡣࠊ766 ࢆྵࡴ㸯NE ௨ୗࡢ࣮ࣝࣉస⏝ࡋ࡚㌿ 㛤ጞ㛵ࢃࡗ࡚࠸ࡿࠋ୍᪉ࠊࢤࣀ࣒ᩓᅾࡍࡿ '63 ࣍ࢵࢺࢫ࣏ࢵࢺࡢస⏝࡛ࡣࠊࢢ࣮ࣟࣂࣝ࡞ࢡ࣐ࣟ ࢳࣥ㧗ḟᵓ㐀ࡢኚࡀ㉳ࡇࡾࠊ⬺จ⦰ࡉࢀࡿ㡿ᇦ ྵࡲࢀࡿ㑇ఏᏊࡢ㌿ࡀάᛶࡉࢀࡿࠋࡇࢀࡣ᪂つ ࡢ㑇ఏᏊⓎ⌧ㄪ⠇ᶵᵓ⪃࠼ࡿࡇࡀ࡛ࡁࡿࠋ 㸲㸧ᮏ◊✲ࢆ㏻ࡌ࡚ࠊࢺ࣏ ,,ș 」ྜయᙧᡂ࡞ࠊ ᐦ᥋࡞㛵ಀࢆᣢࡗ࡚ാࡃࢱࣥࣃࢡ㉁㸦KQ5138 &7&)㸧 ࡀྠᐃࡉࢀࡓࡇࡽࠊ᪂ࡓ࡞ᒎᮃࡀ㛤ࢀࡓࠋ 㸳㸧ࢺ࣏ ,,ș ࡀไᚚࡍࡿ㑇ఏᏊࡣ⮬㛢ࡸ⤫ྜኻ ㄪࡢ㈐௵㑇ఏᏊࡉࢀࡿࡶࡢࡀከ࠸ࠋᚋࠊ &5,635&DV ࡞ࡢࢤࣀ࣒⦅㞟ᢏ⾡ࢆ⏝࠸࡚ࠊࢺ࣏ ,, șࡢ '63 ࣍ࢵࢺࢫ࣏ࢵࢺࢆ᧯సࡍࡿࡇࡼࡾࠊ⢭ ⚄ᝈࡢ⒪ࢆ┠ᣦࡋࡓᛂ⏝◊✲ࡀྍ⬟࡞ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Onoda, A.et al., Nucleic Acids Res. 42, 9005-9020 (2014) Miyaji, M. et al., J. Cell. Biochem. 116, 677-685 (2015) - 437 - බເᨭㄢ㢟㸸2015 ᖺᗘ 㞴ᛶឤᰁࡢཎᅉ࡞ࡿఇṆ⣽⳦ࡢศᏊᶵᵓゎ᫂ ᣮᡓⓗⴌⱆ◊✲ ᖖ⏣ ⪽࣭᪩✄⏣Ꮫ⌮ᕤᏛ⾡㝔 㸺⫼ᬒ࣭┠ⓗ㸼 㑇ఏᏊⓗྠ୍ࡢ༢୍⣽⳦✀ࡢ⣽⳦㞟ᅋ୰࡛࠶ࡗ ࡚ࡶࠊ࠶ࡿ୍ᐃࡢྜ㸦⣙ 10-6㸧࡛ቑṪ㏿ᗘࡀ㏻ᖖ ࡼࡾ㠀ᖖ㐜࠸࠶ࡿ࠸ࡣࡃቑṪࡋ࡞࠸⣽⳦㸦ఇṆ ⣽⳦㸧ࡢள㞟ᅋࡀ⏕ࡌࡿࡇࡀࢃࡗ࡚ࡁࡓࠋᢠ⏕ ≀㉁ࡣ⣽⳦ࡢቑṪࢆᶆⓗࡋ࡚⣽⬊ࢆṚ⮳ࡽࡋࡵ ࡿࠋࡋࡋࠊఇṆ⣽⳦ࡣ⣽⬊ቑṪࢆ⾜ࢃ࡞࠸ࡓࡵࠊ ᢠ⏕≀㉁ࡢస⏝ࢆཷࡅࡎࠊ」ᩘ✀ࡢᢠ⏕≀㉁ᑐࡋ ࡚ᢠᛶࢆ♧ࡋ⏕Ꮡࡋ⥆ࡅࡿࠋఇṆ⣽⳦ࡣቑṪࡢᢚ ไࡍ࡞ࢃࡕ⾲⌧ᆺࡢኚ␗࡛࠶ࡿࡓࡵࠊ㑇ఏᏊᆺࡢኚ ␗࡛࠶ࡿᢠ⏕≀㉁⪏ᛶ⳦ࡣ␗࡞ࡾࠊࡑࡢᛶ㉁ࡣ㠀 㑇ఏᛶ࡛࠶ࡾྍ㏫ⓗ࡛࠶ࡿࠋୖグࡢᛶ㉁ࡽࠊఇṆ ⣽⳦ࡣ⮫ᗋ⌧ሙ࠾࠸࡚ᢠ⏕≀㉁⒪ᚋࡢឤᰁ ⇞ࡢཎᅉ࡞ࡗ࡚࠸ࡿࠋఇṆ⣽⳦ࡣ 1940 ᖺ௦Ⓨぢ ࡉࢀ࡚࠸ࡿࡀࠊឤᰁࡢ⇞ࡢၥ㢟ࡸࠊᛶ㉁ࡢ⏕≀ Ꮫⓗ࡞⯆῝ࡉࡶ㛵ࢃࡽࡎࠊᢠ⏕≀㉁⪏ᛶ⳦ ࡣゎᯒࡀ㐍ࢇ࡛࠸࡞࠸ࠋࡑࡢཎᅉࡢ୍ࡘࡣࠊఇṆ⣽ ⳦ࡢ࣐࣮࣮࢝ࡀᏑᅾࡋ࡞ࡗࡓࡇ࡛࠶ࡗࡓࡀࠊ⏦ ㄳ⪅ࡽࡣ⣽⳦⣽⬊ࡢቑṪࢆᶆⓗࡋࡓୡ⏺ึࡢఇṆ ⣽⳦࣐࣮࣮࢝ᰴࢆ㛤Ⓨࡍࡿࡇᡂຌࡋࡓࠋᮏ◊✲ ࡛ࡣࠊఇṆ⣽⳦ࡢ࣐࣮࣮࢝ᰴࡼࡾఇṆ⣽⳦ࢆศྲྀ ࡋࠊ⥙⨶ⓗ࡞㑇ఏᏊⓎ⌧ゎᯒࢆ⾜࠺ࡇ࡛ఇṆ⣽⳦ ࡢศᏊᶵᵓࡢయീࢆᤊ࠼ࡿࠋ ࡲࡓࠊఇṆ⣽⳦ࡣ⎔ቃࢫࢺࣞࢫࡀຍࢃࡗࡓ㝿ᙧᡂ ࡀಁ㐍ࡉࢀࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࡀࠊຍࢃࡿࢫࢺࣞ ࢫ่⃭ẖ␗࡞ࡿ⤒㊰ࡀࡣࡓࡽ࠸࡚࠸ࡿࡇࡀ♧၀ ࡉࢀ࡚࠸ࡿࠋᚑ᮶ࡢࡼ࠺࡞༢୍ࡢ㑇ఏᏊࢆᶆⓗࡋ ࡓ㑇ఏᏊⓎ⌧ゎᯒ࡛ࡣ⣽⳦ࡢ⤒㊰యࡢẚ㍑ゎᯒࡣ 㞴ࡋࡃࠊලయⓗࡢࡼ࠺࡞㑇ఏᏊࡢⓎ⌧ࡀ␗࡞ࡗ ࡚࠸ࡿࡣ᫂ࡽ࡞ࡗ࡚࠸࡞࠸ࠋᮏ◊✲ㄢ㢟࡛ࡣ 㑇ఏᏊⓎ⌧యീࢆ⥙⨶ⓗẚ㍑ࡍࡿࡇ࡛ࠊ⎔ቃ ࡼࡿ㑇ఏᏊ⤒㊰ࡢ㐪࠸ࡶ᫂ࡽࡍࡿࡇࡀྍ⬟ ࡛࠶ࡾࠊ࡚ࡢ⎔ቃඹ㏻ࡋ࡚Ⓨ⌧ࡍࡿ㑇ఏᏊὀ ┠ࡍࡿࡇ࡛ࠊࡼࡾ㔜せ࡞⸆ࢱ࣮ࢤࢵࢺ࡞ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᣦᑟ ᚑࡗ࡚ࢧࣥ ࣉࣝࢆ ㄪ〇ࡋࡓ ࠋ RNA-sequencing ࡢ⤖ᯝࢆẚ㍑ゎᯒࡍࡿࡇࡼࡗ ࡚ࠊఇṆ⣽⳦ᙧᡂ㛵ࢃࡿ㑇ఏᏊࡀ᫂ࡽ࡞ࡿ ⪃࠼ࡽࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᇵ㣴⎔ቃࢆኚ࠼ࡓ㝿ࠊఇṆ⣽⳦ᙧᡂ㛵ࢃࡿ㑇 ఏᏊࡀ␗࡞ࡗ࡚࠸ࡓሙྜࠊ᪤ ◊✲ࡣࡃ␗࡞ࡿ ᒎᮃࡀᚓࡽࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ≉࡞ࡋࠋ 㸺ᨭࡢෆᐜ㸼 RNA-seq ࢆ⏝࠸࡚ఇṆ⣽⳦ࡢ㑇ఏᏊⓎ⌧ゎᯒࢆࡍ ࡿࡓࡵࡢ᭱㐺࡞᮲௳ࡸࢧࣥࣉࣝㄪᩚ᪉ἲࡢᥦࠋ㠃 ᥋࣓࣮ࣝࡼࡿሗࡽࠊ௨ୗࡢィ⏬ࢆ㐍ࡵ ࡿࡇࢆỴᐃࡋࡓࠋ ࣭RNA-seq (Escherichia coli K12 derivative strain) ࣭ヨᩱᩘ㸸18 (6 samples, 3 biological replicates) ࣭ෆヂ㸸ఇṆ⣽⳦࣐࣮࣮࢝ᰴࢆ 6 ࡘࡢ᮲௳࡛ศྲྀࡋࠊ ࡑࢀࡒࢀ RNA ࢆㄪ〇ࡍࡿࠋ1) ᗣ࡞⣽⳦ࠊ2) ☜⋡ ⓗᙧᡂࡉࢀࡿఇṆ⣽⳦ࠊ3) ᎘Ẽࢫࢺࣞࢫ࡛ᙧᡂࡉ ࢀࡿఇṆ⣽⳦ࠊ4) 㣚㣹≧ែ࡛ᙧᡂࡉࢀࡿఇṆ⣽⳦ࠊ 5) 㓟ᛶࢫࢺࣞࢫ࡛ᙧᡂࡉࢀࡿఇṆ⣽⳦ࠊ6) ࢡ࣒࢜ࣛ ࢭࣥࢩࣥࢢ࡛ᙧᡂࡉࢀࡿఇṆ⣽⳦ ⏦ㄳ⪅ࡢࢧࣥࣉࣝㄪᩚၥ㢟ࡀⓎ⏕ࡋࠊࢧࣥࣉࣝ ࡢᥦ౪ࡀᖜ㐜ࢀ࡚࠾ࡾࠊ1 ᭶ 23 ᪥Ⅼ࡛ゎᯒ ࡣ⮳ࡗ࡚࠸࡞࠸ࠋ1 ᭶ᮎ RNA-sequencing ࢆ⾜࠸ࠊ ⤖ᯝࢆẚ㍑ゎᯒࡍࡿࠋ - 438 - බເᨭㄢ㢟㸸2014ࠥ2015 ᖺᗘ Ꮫಟ㣭ᑟධ siRNA ࡼࡿ㸯㑇ఏᏊ≉␗ⓗࢣ࣒ࣂ࢜ࢣ࣒ᆺ RNA ᖸ΅ἲࡢ㛤Ⓨ ᪂Ꮫ⾡㡿ᇦ◊✲ ⛬ ஂ⨾Ꮚ࣭ᮾிᏛᏛ㝔⌮Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 㻾㻺㻭 ᖸ΅ἲ䛿 㼟㼙㼍㼘㼘㻌㼕㼚㼠㼑㼞㼒㼑㼞㼕㼚㼓㻌㻾㻺㻭㻌㻔㼟㼕㻾㻺㻭㻕䛻䜘䜛⡆౽ 䛺㑇ఏᏊ䝜䝑䜽䝎䜴䞁ἲ䛸䛧䛶ᗈ䛟⏝䛥䜜䛶䛔䜛ᡭἲ䛷 䛒䜚䚸䝀䝜䝮⸆䜔ᚑ᮶䛾㑇ఏᏊ⒪䛻᭰䜟䜛᪂䛧䛔ᡭ ἲ䛸䛧䛶䚸䝀䝜䝮་⛉ᏛⓗほⅬ䛛䜙䛾ᮇᚅ䜒㧗䛔䚹䛧䛛䛧 䛺䛜䜙䚸䛭䛾⮫ᗋᛂ⏝䜈䛾ᐇ⏝䛜䛺䛛䛺䛛ᐇ⌧䛧䛺 䛔⫼ᬒ䛻䛿䚸≉␗ᛶ䜔Ᏻᐃᛶ䛺䛹䛻䛚䛔䛶䚸䜎䛰ゎỴ 䛩䜉䛝㔜せ䛺ၥ㢟䜢ᢪ䛘䛶䛔䜛⌧≧䛜䛒䜛䚹ᡃ䚻䛿䚸䛣 䜜䜎䛷䛻䚸㻾㻺㻭 ᖸ΅䛾ศᏊ䝯䜹䝙䝈䝮ゎᯒ䛾⤖ᯝ䛛䜙䚸 ᭷ຠ䛷ᶆⓗ㑇ఏᏊ≉␗ᛶ䛾㧗䛔 㼟㼕㻾㻺㻭 䛾㓄ิ㑅ᢥἲ 䜢ᵓ⠏䛧䛶䛝䛯䚹䛧䛛䛧䚸䛭䜜䜙䛿ሷᇶ㓄ิୖ䛾ไ㝈䛜 䛝䛔䛯䜑䚸༑ศ䛺ᩘ䛾 㼟㼕㻾㻺㻭 䜢㑅ᢥ䛩䜛䛣䛸䛜㞴䛧䛔䚹 ⛉◊㈝䛾ㄢ㢟䛷䛿䚸ሷᇶ㓄ิ䛻䜘䜛ไ㝈䜢ᅇ㑊䛩䜛䛯 䜑䛾㠀ᖖ䛻᭷⏝䛺ᡭἲ䛸䛧䛶ྛ✀Ꮫಟ㣭䜢⏝䛧䛶䚸 ᶆⓗ㑇ఏᏊ≉␗ⓗ䛺 㼟㼕㻾㻺㻭 䛻䜘䜛 㻾㻺㻭 ᖸ΅ἲ䜢ᵓ⠏䛩 䜛䛣䛸䜢ィ⏬䛧䚸ᐇ䛧䛶䛔䜛䚹㻌 㻌 㻌 䛧䛛䛧䛺䛜䜙䚸ᮏㄢ㢟䛾ᙜึ䛾ணᐃ䜢ୖᅇ䜛㔜せ䛷ᬑ 㐢ᛶ䛾㧗䛔◊✲ᡂᯝ䜢ᚓ䜛䛯䜑䛻䛿䚸䜘䜚ຠ⋡ⓗ䛺 㻾㻺㻭 ᖸ΅ἲ䜢ᵓ⠏䛩䜛ᚲせ䛜䛒䜛䚹≉䛻䚸㻾㻺㻭 ᖸ΅䛻 㛵䜟䜛䛸⪃䛘䜙䜜䜛ᮏ㙐 㻾㻺㻭 ⤖ྜ䝍䞁䝟䜽㉁䛻䛿䚸 㻾㻺㻭 ᖸ΅స⏝䜢᭷ព䛻ಁ㐍䛩䜛䛸⪃䛘䜙䜜䜛䜒䛾䛜䛔䛟 䛴䛛Ꮡᅾ䛩䜛䚹ᮏㄢ㢟䛷䛿䚸䛭䛾䜘䛖䛺ᮏ㙐 㻾㻺㻭 ⤖ ྜ䝍䞁䝟䜽㉁䛾 㼟㼕㻾㻺㻭 䜔 㼙㼕㼏㼞㼛㻾㻺㻭㻌㻔㼙㼕㻾㻺㻭㻕䛸䛾┦స ⏝ 䛻 䛴 䛔 䛶 䚸 䛸 䛧 䛶 㻾㻺㻭㻌 㼕㼙㼙㼡㼚㼛㼜㼞㼑㼏㼕㼜㼕㼠㼍㼠㼕㼛㼚㻌 㻔㻾㻵㻼㻕㻙㻿㼑㼝㼡㼑㼚㼏㼑㻌㻔㻿㼑㼝㻕ἲ䜢⏝䛔䛯ゎᯒ䜢⾜䛖䚹㻌 㸺ᨭࡢෆᐜ㸼㻌 㻾㻺㻭 䝃䜲䝺䞁䝅䞁䜾䜢ไᚚ䛩䜛ᮏ㙐 㻾㻺㻭 ⤖ྜ⺮ⓑ㉁ 㼀㻾㻮㻼 䜢Ⓨ⌧䛧䛶䛔䜛 㻲㼘㼜㻙㻵㼚㻌 㻴㻱㻷㻞㻥㻟 ⣽⬊ᰴ䜢⏝䛔䛯 㻾㻵㻼㻙㻿㼑㼝䠄㻝㻜 ᳨య䠅䜢⾜䛳䛯䚹㻌 㻌 㻌 䛥䜙䛻䚸㻾㻺㻭 䝃䜲䝺䞁䝅䞁䜾䜢ไᚚ䛩䜛ᮏ㙐 㻾㻺㻭 ⤖ ྜ⺮ⓑ㉁ 㼀㻾㻮㻼 䜢Ⓨ⌧䛧䛶䛔䜛 㻲㼘㼜㻙㻵㼚㻌 㻴㻱㻷㻞㻥㻟 ⣽⬊ᰴ 䛚䜘䜃 㻰㼕㼏㼑㼞 䛸⤖ྜ䛷䛝䛺䛔 㼀㻾㻮㻼 Ⓨ⌧ 㻲㼘㼜㻙㻵㼚 ⣽⬊䛷䛾 㻾㻵㻼㻙㻿㼑㼝䠄㻝㻢 ᳨య䠅䚸䛚䜘䜃⣽⬊㉁䌦᰾㛫䜢䝅䝱䝖䝸䞁䜾䛩 䜛 㻾㻺㻭 䝃䜲䝺䞁䝅䞁㛵㐃䝍䞁䝟䜽㉁䛾 㻾㻵㻼㻙㻿㼑㼝䠄㻝㻤 ᳨య䠅 䜢ணᐃ䛧䛶䛔䜛䚹㻌 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㼀㻾㻮㻼 䛿䚸㻴㻵㼂㻙㻝 䛾 㻸㼀㻾 㡿ᇦ䛻Ꮡᅾ䛩䜛䝇䝔䝮䝹䞊䝥≧ 䛾 㼀㻭㻾㻌㻾㻺㻭 䛻⤖ྜ䛩䜛ᅉᏊ䛸䛧䛶ྠᐃ䛥䜜䛯㑇ఏᏊ䛷 䛒䜛䚹୍᪉䛷䚸㼀㻾㻮㻼 䛿ᮏ㙐 㻾㻺㻭 䛸┦స⏝䛩䜛䝗䝯䜲 䞁䛸ᮏ㙐 㻾㻺㻭 ษ᩿㓝⣲䛷䛒䜛 㻰㼕㼏㼑㼞 䛸┦స⏝䛩䜛䝗 䝯䜲䞁䜢䜒䛱䚸㻾㻺㻭 䝃䜲䝺䞁䝅䞁䜾䜢ಁ㐍䛩䜛స⏝䜢䜒䛴 䛣䛸䛜▱䜙䜜䛶䛔䜛䛜䚸䛭䛾ヲ⣽䛺ศᏊᶵᵓ䛿᫂䛷䛒 䛳䛯䚹ᅇ䛾⤖ᯝ䛛䜙䚸䝠䝖䛷䛿 㻞㻘㻜㻜㻜 ⛬ᗘᏑᅾ䛩䜛䛸䛥 䜜䛶䛔䜛 㼙㼕㻾㻺㻭 䛾✀㢮䛻䜘䛳䛶䚸㼀㻾㻮㻼 䛜ዲ䜣䛷⤖ྜ䛩 䜛䜒䛾䛸⤖ྜ䛧䛻䛟䛔䜒䛾䛜䛒䜛䛣䛸䛜᫂䜙䛛䛻䛺䛳䛯䚹䛥 䜙䛻 㼙㼕㻾㻺㻭 䛾✀㢮䛻䜘䛳䛶䛿䚸㼀㻾㻮㻼 䛸┦స⏝䛧䛶䛔 䜛 㻰㼕㼏㼑㼞 䛻䜘䜛ษ᩿䛜 㼙㼕㻾㻺㻭 ᮏ㙐䛾∦ഃ䛾 㻾㻺㻭 㙐䛷 㜼ᐖ䛥䜜䜛䜘䛖䛺ᶵᵓ䛜䛒䜛ྍ⬟ᛶ䜒᫂䜙䛛䛻䛺䛳䛯䚹䜎 䛯䚸䛣䜜䜎䛷䛿 㻭㼓㼛 䝍䞁䝟䜽㉁䛾⾲㠃䛻㍕䛳䛶䛔䜛ᡂ⇍ ᆺ 㼙㼕㻾㻺㻭 䛜ᶆⓗ䛸䛩䜛 㼙㻾㻺㻭 䛸⤖ྜ䛩䜛䛸⪃䛘䜙䜜䛶䛔 䛯䛜䚸ᅇ䛾ゎᯒ䛛䜙 㼀㻾㻮㻼 䜒 㼙㼕㻾㻺㻭 䛸 㼙㻾㻺㻭 䛷ᙧᡂ䛥 䜜䜛ᮏ㙐 㻾㻺㻭 䛻⤖ྜ䛧䛶䛔䜛ྍ⬟ᛶ䛜♧၀䛥䜜䛯䚹 䛣䛾䜘䛖䛺ᮏ㙐 㻾㻺㻭 䛻 㼀㻾㻮㻼 䛜⤖ྜ䛩䜛ྍ⬟ᛶ䛿༑ ศ⪃䛘䜙䜜䜛䛯䜑䚸㼀㻾㻮㻼 䛿 㼙㼕㻾㻺㻭 䛸 㼙㻾㻺㻭 䛾⤖ྜ䜢⿵ ᙉ䛩䜛䜘䛖䛻ാ䛟ሙྜ䛜䛒䜛䛾䛛䜒䛧䜜䛺䛔䚹䛣䛾䜘䛖䛺 Ⅼ䜢᫂䜙䛛䛻䛩䜛䛯䜑䛻䚸㔝⏕ᆺ 㼀㻾㻮㻼 䛸┦స⏝䛩䜛 㛗䛔 㻾㻺㻭 䛾ゎᯒ䜒㐍䜑䛶䛔䜛䚹㻌 㻌 㻌 䛥䜙䛻䚸㼀㻾㻮㻼 䛿䠏䛴䛾ᮏ㙐 㻾㻺㻭 ⤖ྜ䝗䝯䜲䞁䜢䜒䛴 䛜䚸䜟䜜䜟䜜䛿䚸㼀㻾㻮㻼 䛜䠎䛴┠䛸䠏䛴┠䛾 㼐㼟㻾㻮㻰 䛾㛫 䛷ษ᩿䛥䜜䜛䝍䞁䝟䜽㉁䛷䛒䜛䛣䛸䜢ぢฟ䛧䛯䚹ษ᩿Ⅼ䛿 䛱䜗䛖䛹䠍䚸䠎␒┠䛾ᮏ㙐 㻾㻺㻭 ┦స⏝䝗䝯䜲䞁䛸䠏␒ ┠䛾䝍䞁䝟䜽㉁┦స⏝䝗䝯䜲䞁䜢ศ᩿䛩䜛⨨䛻䛒䜚䚸 㼀㻾㻮㻼 䛿ษ᩿䛥䜜䜛䛣䛸䛻䜘䜚ᶵ⬟ⓗ䛻䛝䛟ኚ䛩䜛䛣 䛸䛜ᮇᚅ䛥䜜䜛䚹䛭䛣䛷䚸≉䛻 㼀㻾㻮㻼 䛾ᵓ㐀ኚ䛜䛹䛾䜘 䛖䛺ᶵ⬟䛾ኚ䜢䜒䛯䜙䛩䛾䛛䛸䛔䛖Ⅼ䛻䛴䛔䛶ゎ᫂䛩 䜛䛯䜑䛻䚸㔝⏕ᆺ䛸䚸䠏␒┠䛾䝗䝯䜲䞁䛜Ḟᦆ䛧䚸䠍␒┠䛸 䠎␒┠䛾䝗䝯䜲䞁䛾䜏䜢䜒䛴ษ᩿ᆺ 㼀㻾㻮㻼 䛾୧⪅䛻ὀ┠ 䛧䛯ゎᯒ䜢⾜䛳䛯䚹䛭䛾⤖ᯝ䚸䠏␒┠䛾䝗䝯䜲䞁䜢Ḟኻ䛧 䛯 㼀㻾㻮㻼 䛿 㼙㼕㻾㻺㻭 䛾䝥䝻䝉䝅䞁䜾䛜䛷䛝䛺䛔䛣䛸䛜᫂䜙 䛛䛻䛺䛳䛯䚹㼀㻾㻮㻼 䛿䠏␒┠䛾䝗䝯䜲䞁䛜Ḟኻ䛩䜛䛸䚸 㻰㼕㼏㼑㼞 䛸䛾┦స⏝䛜䛷䛝䛺䛔䛯䜑䛻 㼙㼕㻾㻺㻭 䛾䝥䝻䝉䝅 䞁䜾䛜䛷䛝䛺䛔ᙧ䛻ᶵ⬟㌿䛩䜛䛣䛸䛜♧၀䛥䜜䛯䚹䛣 䛾䛣䛸䜢᫂䜙䛛䛻䛩䜛䛯䜑䛻䚸㻰㼕㼏㼑㼞 䛸䛾┦స⏝䛜䛷䛝 䛺䛔 㼀㻾㻮㻼 䛾 㻲㼘㼜㻙㻵㼚 ⣽⬊䜢ᵓ⠏䛧䚸䛭䛾 㻾㻵㻼㻙㻿㼑㼝 䜢⾜䛖 ணᐃ䛷䛒䜛䚹㻌 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡇࢀࡲ࡛ࡢᨭࡼࡾࠊRNA ࢧࣞࣥࢩࣥࢢ㛵ࢃ ࡿᮏ㙐 RNA ⤖ྜࢱࣥࣃࢡ㉁ TRBP ࡢ᪂ࡓ࡞స⏝ ᶵᗎࡀ᫂ࡽ࡞ࡗ࡚ࡁࡓࠋࡲࡓࠊ⌧ᅾࠊRNA ࢧ ࣞࣥࢩࣥࢢ㛵ࢃࡿࡶ࠺㸯ࡘࡢᮏ㙐 RNA ⤖ྜࢱ ࣃࢡ㉁࡛࠶ࡿ TNRC6A ࡘ࠸࡚ࡶྠᵝࠊRIP-Seq ࢆ⏝࠸࡚ࠊmiRNA ࡸ siRNAࠊ࠾ࡼࡧࡑࢀࡽࡢᶆⓗ mRNA ࡢ┦స⏝ࢆ᫂ࡽࡍࡿணᐃ࡛࠶ࡿࠋࡇ ࡢࡼ࠺࡞ヲ⣽࡞ゎᯒࡼࡾࠊࡇࢀࡲ࡛࡛᫂࠶ࡗࡓ RNA ࢧࣞࣥࢩࣥࢢࡢ᪂ࡓ࡞ศᏊᶵᵓࢆ᫂ࡽ ࡋࠊࡼࡾຠ⋡ⓗ࡞ RNA ࢧࣞࣥࢩࣥࢢㄏᑟᶵᵓࡢ᳨ ウࢆ⾜࠺ணᐃ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᨭࡢᡂᯝࢆබ⾲ࡋࡓ㞧ㄅㄽᩥࡣࡲࡔ࡞࠸ࡀࠊ⌧ᅾ ‽ഛ୰࡛࠶ࡿࠋ - 439 - බເᨭㄢ㢟㸸2014ࠥ2015 ᖺᗘ GVHD ࡼࡿ࣓࣍࢜ࢫࢱࢩࢫ◚⥢ࡢ࣓࢝ࢽࢬ࣒ ᇶ┙◊✲㸦㹀㸧 ㇏ᔱ ᓫᚨ࣭ᾏ㐨ᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ㸼 ྠ ✀ 㐀 ⾑ ᖿ ⣽ ⬊ ⛣ ᳜ (allogeneic hematopoietic stem cell transplantation: allo-HSCT)ࡣ, 㞴ᛶⓑ⾑➼ᑐࡋ࡚၏୍ࡢ᰿ⓗ⒪ἲ࡞ࡿࠋ ࡋࡋ, ⛣᳜ᚋࡣ⛣᳜∦ᑐᐟ(graft-versus-host disease: GVHD)࣭ឤᰁ➼ࡢ⮴ⓗ࡞ྜేࡀၥ㢟 ࡞ࡿࠋGVHD ࡣࢻࢼ࣮⏤᮶ࡢ T ⣽⬊ࡀ, ࣞࢩࣆ࢚ࣥ ࢺࡢࣟᢠཎࡼࡗ࡚άᛶࡉࢀ, ㌟ࡢ⮚ჾ㞀ᐖ ࢆ㉳ࡇࡍ་ཎᛶࡢᝈ࡛࠶ࡾ, ࡑࡢண㜵ࡣ, Ᏻ ⛣᳜ࢆ⾜࠺ࡓࡵᚲ㡲࡛࠶ࡿࠋ⌧⾜ࡢ GVHD ண㜵ࡣ, ࢝ࣝࢩࢽ࣮ࣗࣜࣥ㜼ᐖ➼ࡼࡿᙉຊ࡞චᢚไ ࡼࡗ࡚⾜ࢃࢀ࡚࠾ࡾ, ⛣᳜ࡢ┠ⓗ࡛࠶ࡿᢠ⭘⒆ຠ ᯝࢆᡴࡕᾘࡋ, ᪥ぢឤᰁࡢⓎࢆᣍࡃྍ⬟ᛶࡀ ࠶ࡿࠋallo-HSCT ࢆᏳ⾜ࡍࡿࡓࡵࡣ, GVHD ࡢែ⏕⌮ࢆヲ⣽᳨ウࡋ, ᪂ࡓ࡞ GVHD ண㜵ἲ࣭ ⒪ἲࢆ㛤Ⓨࡍࡿᚲせࡀ࠶ࡿࠋ ⏦ㄳ⪅ࡣ᭱㏆, ⭠⟶ࡢ Paneth ⣽⬊ࡀ GVHD ࡢᶆⓗ࡞ࡿࡇࢆⓎぢࡋࡓ(Eriguchi et al, Blood 2012)ࠋPaneth ⣽⬊ࡣ, ᢠ⳦࣌ࣉࢳࢻ࡛࠶ࡿD-defensin ➼ࢆ⏘⏕ࡍࡿࡇࡼࡗ࡚, ཎᛶ⣽⳦ࡢቑṪࢆᢚ ไࡋ, ⭠⟶ࡢᖖᅾ⣽⳦ྀࢆಖࡗ࡚࠸ࡿࠋGVHD ࡀⓎ ࡍࡿ, Paneth ⣽⬊ࡣ㏿ࡸῶᑡࡋᢠ⳦࣌ࣉࢳ ࢻࡢ⏘⏕ࡀపୗࡋ, ཎᛶ⣽⳦ࡢⴭ᫂࡞ቑṪࡼࡗ ࡚⭠ෆ⣽⳦ྀࡢ␗ᖖ(dysbiosis)ࡀ⏕ࡎࡿࠋDysbiosis ࡣ, ᭦࡞ࡿ GVHD ࡢᝏࡸᩋ⾑ࡢⓎࢆᣍࡃࠋࡇ࠺ࡋ ࡓ͆GVHD ឤᰁࡢࢡࣟࢫࢺ࣮ࢡ͇ࢆ᩿ࡕษࡿࡇ ࡣ, ᗈỗ࡞චᢚไࢆకࢃ࡞࠸Ᏻࡘ᭷ຠ࡞ḟ ୡ௦ࡢ GVHD ண㜵࣭⒪ἲ࡞ࡿྍ⬟ᛶࡀ࠶ࡿࠋ 㸺┠ⓗ㸼 ᡃࠎࡣ, ⭠⟶ୖ⓶ቑṪᅉᏊ X ࢆᢞࡍࡿࡇ ࡼࡾ࣐࢘ࢫࡢ Paneth ⣽⬊ࡀⴭ᫂ቑຍࡍࡿࢆ ぢ࠸ࡔࡋࡓࠋࡇࡢ Paneth ⣽⬊ࡢቑຍࡣ, ⭠⟶ෆࡢᢠ ⳦࣌ࣉࢳࢻ⃰ᗘࡢⴭ᫂࡞ୖ᪼ࢆࡲࡡࡁ, ⭠⟶ෆᚤᑠ ⎔ቃⓗ࡞ኚࡀ⏕ࡌࡉࡏࡿࢆぢ࠸ࡔࡋࡓࠋࡉ ࡽ, ⭠⟶ୖ⓶ቑṪᅉᏊ X ࢆ⛣᳜๓ᚋᢞࡍࡿ, GVHD ࡀᢚไࡉࢀࡓࡀ, ࡇࢀࡣ dysbiosis ࡢᢚไࡼ ࡿࡢ࡛ࡣ࡞࠸ࡢ௬ㄝ⮳ࡗࡓࠋGVHD ⭠ෆ⣽ ⳦ྀࡢኚࡀᐦ᥋⤖ࡧࡘ࠸࡚࠸ࡿࡣ, ᡃࠎࡢࢢ ࣮ࣝࣉࡸࡢࢢ࣮ࣝࣉࡽ᪤Ⓨ⾲ࡉࢀ࡚࠾ࡾ,⭠ෆ ⣽⳦ྀࢆ⭠⟶ୖ⓶ቑṪᅉᏊ X ࡼࡗ࡚ࢥࣥࢺ࣮ࣟࣝ ࡍࡿࡇࡼࡾ, GVHD ࢆᢚไࡍࡿࡇࡀ࡛ࡁࢀࡤ, ࡇࢀࡣ᪂ࡋ࠸⛣᳜ἲࡢ㛤Ⓨࡢࡘ࡞ࡀࡿ⪃࠼ࡽࢀ ࡿࠋࡇ࠺ࡋࡓ௬ㄝࢆド᫂ࡍࡿࡓࡵ, ⛣᳜๓ᚋ⭠⟶ୖ ⓶ቑṪᅉᏊ X ࢆᢞࡋ, ⛣᳜ᚋࡢ⭠ෆ⣽⳦ྀኚࢆ 16S rRNA 㡿ᇦࡢࢩ࣮ࢣࣥࢫࡼࡗ࡚ホ౯ࡍࡿࡇ ࡋࡓࠋ ᡃࠎࡣ, ࡉࡽ⭠ෆ⣽⳦ྀࡢኚࡀ, ⛣᳜ ᚋࡢ⏕╔ᙳ㡪ࢆཬࡰࡍࢆぢ࠸ࡔࡋ࡚࠸ࡿࠋ⛣᳜ ᚋࡢ⏕╔ࡢࢫࣆ࣮ࢻࡸ⏕╔⋡ࡣ, ⛣᳜ࡢᡂྰࢆỴᐃ ࡙ࡅࡿ㔜せ࡞せ⣲࡛࠶ࡿࠋ⛣᳜ᚋࡢ⭠ෆ⣽⳦ྀࡢኚ , ⏕╔ࡢ㛵ಀࢆࡉࡽヲ⣽᳨ウࡍࡿࡓࡵ, 16S rRNA 㡿ᇦࡢࢩ࣮ࢣࣥࢫࡼࡗ࡚, ⏕╔࣭ᣄ⤯ ⭠ෆ⣽⳦ྀࡢ㛵㐃ࢆゎ᫂ࡋࡓ࠸ࠋ ᭱㏆, ⭠ෆ⣽⳦ྀࡢኚࡀ┿⳦ྀࡢኚ ࡶࡘ࡞ࡀࡿࡀ♧ࡉࢀࡓ(Kim et al, Cell Host Microbe. 2014)ࠋࡇࢀࡲ࡛ᡃࠎࡣ㐀⾑ᖿ⣽⬊⛣᳜ᚋ, ⭠ෆ⣽ ⳦ྀࢆࢥࣥࢺ࣮ࣟࣝࡍࡿࡇࡼࡗ࡚⛣᳜ࡢᡂ⦼ࢆ ྥୖࡉࡏࡼ࠺⪃࠼࡚ࡁࡓࠋࡋࡋ, 㐀⾑ᖿ⣽⬊⛣᳜ ࡢ㌿ᖐࡣ, ⣽⳦ྀࡔࡅ࡛ࡣ࡞ࡃ┿⳦ྀࡢኚࡀ㛵 ࡋ࡚࠸ࡿྍ⬟ᛶࡶ㧗࠸ࠋᡃࠎࡣ᪤┿⳦ࡢ⳦యᡂ ศࡼࡗ࡚⫵≉␗ⓗ࡞ GVHD ࡀⓎࡍࡿࢆࡘࡁ ࡵ࡚࠸ࡿ (Uryu et al, Blood 2015)ࠋᡃࠎࡣ, ࡲࡎ㐀 ⾑ᖿ⣽⬊⛣᳜๓ᚋࡸ GVHD ࡢⓎ๓ᚋ࡛⭠ෆ┿⳦ྀ ࡢኚࡀ⏕ࡌ࡚࠸ࡿ࠺ࢆ, 18S ITS 㡿ᇦࡢࢩ࣮ ࢣࣥࢫࡼࡗ࡚ゎᯒࡋࡓ࠸ࠋ 㸺ᨭࡢෆᐜ㸼 1. ⛣᳜๓ᚋᵝࠎ࡞ฎ⨨ࢆࡋࡓ࣐࢘ࢫࡢ⣅౽ࡽ DNA ࢆᢳฟࡋ, 16S rRNA 㡿ᇦࢆቑᖜࡋࡓ PCR ⏘≀ࡢ㓄ิࢆ MiSeq ࢆ⏝ࡋ࡚ㄞࢇ࡛㡬࠸ࡓࠋ ࡇࢀࡽࡢ㓄ิࢆゎᯒࡋ࡚㡬ࡁ, ⭠ෆ⣽⳦ྀࢆᵓ ᡂࡍࡿ⳦ࡑࡢቑῶ➼ࢆゎ᫂ࡋ࡚㡬࠸ࡓࠋ 2. ⛣᳜๓ᚋ࡛ᵝࠎ࡞ฎ⨨ࢆࡋࡓ࣐࢘ࢫࡢ⣅౽ࡽ DNA ࢆᢳฟࡋ, 18S ITS 㡿ᇦࢆቑᖜࡋࡓ PCR ⏘ ≀ࡢ㓄ิࢆ MiSeq ࢆ⏝ࡋ࡚ㄞࢇ࡛㡬࠸ࡓࠋࡇ ࢀࡽࡢ㓄ิࢆゎᯒࡋ࡚㡬ࡁ, ⭠ෆ┿⳦ྀࢆᵓᡂ ࡍࡿ┿⳦ࡑࡢቑῶ➼ࢆゎ᫂ࡋ࡚㡬࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 1. ⭠⟶ୖ⓶ቑṪᅉᏊ X ࡣ Paneth ⣽⬊ࢆቑṪࡉࡏ, ⭠⟶ࡢD-defensin ศἪࢆቑຍࡉࡏࡿ: ⭠⟶ୖ⓶ ቑṪᅉᏊ X ትnaïve ࣐࢘ࢫ 6 ᪥㛫ᢞࡍࡿ, ༑ᣦ⭠ࡽᅇ⭠ࡲ࡛ࡢ࡚ࡢᑠ⭠㡿ᇦ࡛ Paneth ⣽⬊ᩘࡀ᭷ពቑຍࡋࡓࠋPaneth ⣽⬊ࡀ⏘ ⏕ࡍࡿᢠ⳦࣌ࣉࢳࢻ࡛࠶ࡿD-defensin ࡢ⃰ᗘࢆ⣅ ౽୰࡛ ᐃࡋࡓࡇࢁ㸪D-defensin ⃰ᗘࡣ, ⭠⟶ ୖ⓶ቑṪᅉᏊ X ࢆᢞࡋࡓ࣐࢘ࢫ࡛᭷ពୖ᪼ ࡋ࡚࠸ࡓࠋḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ࢆ⏝ࡋࡓ⭠ෆ⣽ ⳦ྀࡢゎᯒࡼࡗ࡚, ⭠⟶ୖ⓶ቑṪᅉᏊ X ࢆᢞ ࡉࢀࡓ naïve ࣐࢘ࢫࡢ⣅౽࡛ࡣ⭠ෆ⣽⳦ྀࡢ᭷ ព࡞ᵓᡂኚࢆㄆࡵࡽࢀ࡞࠸ࡇࡀุ᫂ࡋࡓࠋࡇ ࢀࡣ Naïve ࣐࢘ࢫࡢ⭠ෆᖖᅾ⳦ࡣD-denfensin 㧗ᗘ࡞⪏ᛶࢆᣢࡘࡓࡵ⪃࠼ࡽࢀࡿࠋ 2. ⭠⟶ୖ⓶ቑṪᅉᏊ X ࡢᢞࡼࡗ࡚, GVHD ࡼࡿD-defensin ࡢศἪയᐖࡀ㍍ῶࡉࢀࡿ: ࣐࢘ࢫ ࡢྠ✀㦵㧊⛣᳜ᚋ, ⛣᳜ᚋ 5 ᪥┠ࡣࢻࢼ࣮T ⣽ ⬊ࡀ⭠⟶⢓⭷ᾐ₶ࡋ, GVHD ࡀⓎࡍࡿࠋ GVHD ࢆⓎࡋࡓ࣐࢘ࢫ࡛ࡣ Paneth ⣽⬊ࡀ ࢇᾘኻࡍࡿࡀ, ⭠⟶ୖ⓶ቑṪᅉᏊ X ࢆᢞࡍ ࡿ⛣᳜ᚋ 4 ᪥┠ 7 ᪥┠ࡢ Paneth ⣽⬊ᩘࡣ㸪 GVHD ࢆⓎࡍࡿࢥࣥࢺ࣮࣐ࣟࣝ࢘ࢫẚ㍑ࡋ ࡚᭷ពከࡗࡓࠋGVHD ࣐࢘ࢫ࠾ࡅࡿ Paneth ⣽⬊ࡢῶᑡࡣ, ⣅౽୰ࡢD-defensin ࡢᾘኻࢆᣍ࠸ ࡓࠋ⭠⟶ୖ⓶ቑṪᅉᏊ X ࡼࡿ Paneth ⣽⬊ಖㆤ ࡼࡗ࡚, ⛣᳜ᚋ 7 ᪥┠ࡢ⣅౽୰ࡢD-defensin ⃰ - 440 - 3. 4. 5. 6. ᗘࡀ᭷ពୖ᪼ࡋ࡚࠸ࡓࠋ ⭠⟶ୖ⓶ቑṪᅉᏊ X ࡢᢞࡼࡗ࡚, GVHD ࡼࡿ dysbiosis ࡀண㜵ࡉࢀ GVHD ࡀ㍍ῶࡉࢀࡿ: ⛣᳜ᚋ 7 ᪥┠᥇ྲྀࡋࡓ⣅౽ࡽ DNA ࢆᢳฟࡋ, PCR ἲࡼࡗ࡚ 16S rRNA 㡿ᇦࢆቑᖜࡋࡓᚋ, ḟୡ௦ࢩ࣮ࢣࣥࢧ࣮MiSeq ࢆ⏝ࡋ࡚㓄ิࢆゎ ㄞࡋࡓࠋࢥࣥࢺ࣮ࣟࣝࡋ࡚, GVHD ࢆⓎࡋ࡞ ࠸, ྠ⣔(Syngeneic)⛣᳜ࢆ⾜ࡗࡓ࣐࢘ࢫࡢ⣅౽ ࡶゎᯒࡋࡓࠋࡇࢀࡲ࡛ࡢ◊✲ࡼࡗ࡚, GVHD ⩌ ࡛ࡣ Syngeneic ⩌ẚ㍑ࡋ࡚ dysbiosis ࡢࡓࡵ⣽ ⳦ࡢከᵝᛶ(diversity)ࡀపୗࡍࡿࡇࡀ▱ࡽࢀ࡚ ࠸ࡓࡀ, ᮏ◊✲࡛ࡶ Shannon ᣦᩘࡸ Simpson ᣦ ᩘ࡞ࡢከᵝᗘᣦᩘࢆ᳨ウࡋࡓࡇࢁ diversity ࡢపୗࡀぢࡽࢀࡓࠋࡲࡓ, GVHD ⩌࡛ࡣ Syngeneic ⩌ẚ㍑ࡋ࡚ Firmicutes 㛛࡞ࡢᖖᅾ⣽⳦ྀࢆ ᙧᡂࡍࡿ⣽⳦ࡀῶᑡࡋ, Escherichia/Shigella ➼ࡢ ཎᛶ⣽⳦ࡢⴭ᫂࡞ቑṪࡀぢࡽࢀࡓࠋ㦫ࡃࡁࡇ ࡇ࠺ࡋࡓ GVHD ࡛ぢࡽࢀࡓ dysbiosis ࡣ, ⭠ ⟶ୖ⓶ቑṪᅉᏊ X ࡢᢞࡼࡗ࡚ࢇ ᢚไࡉࢀ, ⳦ࡢከᵝᛶࡶᅇࡋ࡚࠸ࡓࠋ PERMANOVA ゎᯒࢆ⏝࠸ࡿ, GVHD ⩌ࡢ⳦ྀ ࡣ naïve ࣐࢘ࢫࡸ syngeneic ⩌ࡢ⳦ྀ᭷ព␗ ࡞ࡗ࡚࠸ࡓࡀ, ྠ✀㦵㧊⛣᳜ᚋ⭠⟶ୖ⓶ቑṪ ᅉᏊ X ࢆᢞࡋࡓ࣐࢘ࢫࡢ⳦ྀࡣ naïve ࣐࢘ࢫࡸ syngeneic ⩌᭷ព࡞ᕪࢆᣢࡓ࡞ࡗࡓࠋࡇ࠺ࡋ ࡓ dysbiosis ࡢᢚไࡣ, ⛣᳜ᚋࡢ donor T ⣽⬊ࡢ⭠ ⟶ࡢᾐ₶ࢆᢚไࡋ, GVHD ࢆ㍍ῶࡋ⏕Ꮡ⋡ࡶᨵ ၿࡋ࡚࠸ࡓࠋ ᢠ⳦࣌ࣉࢳࢻࡢᢞ࡛⛣᳜ᚋࡢ dysbiosis ࡀᢚไ ࡛ࡁࡿ: ⛣᳜ᚋᢠ⳦࣌ࣉࢳࢻࢆ⤒ཱྀᢞࡋ, dysbiosis ࡀᢚไࡉࢀࡿྰࢆ⌧ᅾゎᯒ୰࡛࠶ ࡿࠋ ⭠ෆ⣽⳦ྀࡣ⛣᳜ᚋࡢ⏕╔㛵ࡍࡿ: ⛣᳜ᚋ ࡢዲ୰⌫ῶᑡᮇࡣ, ෆᅉᛶࡢ㐀⾑ᅉᏊࡢ⏘⏕ ࡀቑຍࡋ࡚⏕╔ࢆಁ㐍ࡋ࡚࠸ࡿࡀ, ⭠ෆẅ⳦ࢆ ⾜࠺, 㐀⾑ᅉᏊࡢ⏘⏕ஹ㐍ࡀࡳࡽࢀ࡞ࡃ࡞ࡾ, ⏕╔ࡀ㐜ᘏࡍࡿࢆⓎぢࡋࡓࠋ⌧ᅾ, ⏕╔㛵 ࡍࡿ⣽⳦ࡢ✀㢮ࢆ✺ࡁṆࡵࡿࡓࡵ, ࣞࢩࣆ࢚ ࣥࢺ࣐࢘ࢫࡢ⣅౽ࡽᢳฟࡋࡓ DNA ࡢ, 16S rRNA 㡿ᇦࡢゎᯒࢆ⾜ࡗ࡚࠸ࡿࡇࢁ࡛࠶ࡿࠋ ┿⳦ྀࡢኚ: ࣐࢘ࢫࡢྠ✀࠾ࡼࡧྠ⣔⛣᳜ᚋ ࡢ⣅౽ࢆ᥇ྲྀࡋ, ┿⳦ᒙࡢኚࢆゎᯒࡋ࡚࠸ࡿ ࡇࢁ࡛࠶ࡿ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ◊✲ࡼࡗ࡚, ⭠ෆ⣽⳦ྀࢆࢥࣥࢺ࣮ࣟࣝࡍࡿࡇ ࡀ, GVHD ᢚไࡘ࡞ࡀࡿࡇࡀ♧ࡉࢀࡓࠋࡇࢀ ࡲ࡛⮫ᗋࡢ⌧ሙ࠾࠸࡚, ᢠ⏕ࡼࡿ⭠ෆẅ⳦ ࡼࡗ࡚ GVHD ࡢண㜵ࢆ⾜ࡗ࡚࠸ࡿࡀ, ᢠ⏕࡛ࡢ⭠ ෆ⣽⳦ྀࡢࢥࣥࢺ࣮ࣟࣝࡣ࠸ࡃࡘࡢḞⅬࡀᣦ ࡉࢀ࡚࠸ࡿࠋ⭠ෆࡢᖖᅾ⳦ࡣไᚚᛶ T ⣽⬊ࡢㄏᑟ ࡼࡗ࡚, ᭷ᐖ࡞චᛂࢆᢚไࡋ࡚࠸ࡿࡇࡀ▱ࡽ ࢀ࡚࠸ࡿࡀ, ᢠ⏕ࡣࡇ࠺ࡋࡓ᭷┈࡞ᖖᅾ⳦ࡶẅ⳦ ࡋ࡚ࡋࡲ࠺ࠋࡲࡓ, ࡇ࠺ࡋࡓᗈỗ࡞ᢠ⏕ࡢẅ⳦ຠᯝ ࡣ, ⪏ᛶ⳦ࡢ⦾Ṫࢆᣍ࠸࡚ࡋࡲ࠺ࠋᡃࠎࡢ᪉ἲ࡛ࡣ, ⭠⟶ୖ⓶ቑṪᅉᏊ X ࡼࡿ Paneth ⣽⬊ࡢቑᡂࡼࡿ D-defensin ⏘⏕ࡢಁ㐍, ࡲࡓࡣᢠ⳦࣌ࣉࢳࢻ࡛࠶ࡿ D-defensin ࢆ┤᥋ᢞࡍࡿࡇࡼࡗ࡚, GVHD ࡢ㍍ ῶࢆࡣࡗ࡚࠸ࡿࠋD-defensin ࡣ, ཎᛶ⣽⳦ࡣᙉ ຊ࡞ẅ⳦ຠᯝࢆᣢࡘ୍᪉࡛, ᖖᅾ⳦యࡍࡿẅ⳦ຠ ᯝࡣᙅ࠸ࡇࡀ▱ࡽࢀ࡚࠸ࡿ (Masuda et al. J Innate Immun 2011)ࠋࡑࡢࡓࡵ, ᭷┈࡞ᖖᅾ⳦ࢆಖࡕ, ཎ ᛶ⣽⳦ࡢቑᡂࢆᢚไࡍࡿ࠸࠺, ⌮ⓗ࡞ GVHD ண 㜵ἲ࡞ࡿྍ⬟ᛶࡀ࠶ࡿࠋࡇ࠺ࡋࡓ᪂ࡋ࠸, ⭠ෆ⣽⳦ ࡢࢥࣥࢺ࣮ࣟࣝἲࡣ, ⛣᳜ࡢࡳ࡛ࡣ࡞ࡃ, ᵝࠎ࡞ ᝈ᭷⏝࡛࠶ࡿྍ⬟ᛶࡀ࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 [ㄽᩥ] ⌧ᅾᢞ✏‽ഛ୰ [Ꮫ➼Ⓨ⾲] 1. Teshima T. Protection of the tissue stem cell and niche system as a novel approach to 2nd Australia-Japan control GVHD, Haematology Consortium, Sep 11-12 2015, Brisbane, Australia. 2. Hayase E, et al. R-Spondin1 promotes Paneth cell growth, maintains intestinal microbial ecology, and ameliorates graft-versus-host disease, 57th ASH Annual Meeting, Dec 6th 2015, Orlando, FL, USA. 3. Teshima T. Emerging concepts on tissue injury in GVHD. BMT Tandem Meeting 2016, Feb 18th 2016, Honolulu, HI, USA. (Ⓨ⾲ண ᐃ) - 441 - බເᨭㄢ㢟㸸2010 ᖺᗘ ࣓ࢲ࢝㏫㑇ఏᏛⓗᡭἲࢆᇶ┙ࡋࡓಶయ࣭⤌⧊࡛ࣞ࣋ࣝࡢᦆയᛂ⟅ゎᯒ⣔ࡢ☜❧ ᇶ┙◊✲㸦㹑㸧 ⸨ᇽ ๛࣭㜰ᏛᏛ㝔་Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 㑇ఏᏊࡢᶵ⬟ゎᯒ◊✲࠾࠸࡚ࠊ✺↛ኚ␗యࡣᴟ ࡵ࡚᭷⏝࡞ሗࢆᥦ౪ࡍࡿࠋࡋࡋ࡞ࡀࡽࠊ┠ⓗ ࡍࡿ㑇ఏᏊࡢኚ␗యࢆᚓࡿ㏫㑇ఏᏛⓗᡭἲࡣ୍㒊ࡢ ᐇ㦂ࣔࢹࣝ⏕≀࡛ࡋ⏝࡛ࡁ࡞ࡗࡓࠋࡑࡇ࡛ࠊ ᙜ ◊ ✲ ᐊ ࡛ ࡣ ࠊ 7,//,1*7DUJHWHG ,QGXFHG /RFDO /HVLRQV,1*HQRPHἲࢆ⏝࠸࣓ࢲ࢝࠾࠸࡚㏫㑇ఏ Ꮫⓗᡭἲࡢ☜❧ࢆ┠ᣦࡋ࡚ࡁࡓࠋ7,//,1* ἲࡣࠊձ 㞝ぶࢆᏛኚ␗ཎฎ⌮ࡋࡓᚋࠊṇᖖ㞤㓄ࡋࠊࡑ ࡢ ) ಶయࡢࢤࣀ࣒ '1$ ࠾ࡼࡧ⢭Ꮚࢆ⤖ಖᏑࡋࠊኚ ␗ᑟධಶయࢤࣀ࣒ '1$ ࣛࣈ࣮ࣛࣜࢆసᡂࡍࡿࠊղ ࢤࣀ࣒ '1$ ࣛࣈ࣮ࣛࣜࢆ⏝࠸ࠊ┠ⓗ㑇ఏᏊኚ␗ ࡀᑟධࡉࢀࡓಶయࢆࢫࢡ࣮ࣜࢽࣥࢢࡍࡿࠊճኚ␗ࡀ ྠᐃࡉࢀࢀࡤࠊ⤖⢭Ꮚࣛࣈ࣮ࣛࣜࡽ )ಶయࢆ సᡂࡋࠊ࣊ࢸࣟኚ␗ ) ಶయྠኈࡢࡅྜࢃࡏࡼࡾ ) ࠾࠸࡚ኚ␗࣍ࣔಶయࢆᚓࡿࠊ࠸࠺୍㐃ࡢᡭἲ ࡛࠶ࡿࠋࢫࢸࢵࣉ㸰࠾࠸࡚ࡣࠊຠ⋡ⓗ 613V㸦ㄏ Ⓨኚ␗㸧ྠᐃࢆ⾜࠼ࡿ +50+LJK5HVROXWLRQ0HOWLQJ ἲࢆ㛤Ⓨࡋࠊࣛࣈ࣮ࣛࣜࡢࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡗ ࡚ࡁࡓࠋࡋࡋ࡞ࡀࡽࠊసᡂࣛࣈ࣮ࣛࣜࢆࡼࡾຠ ⋡ⓗ⏝ࡍࡿࡣࠊࡼࡾࣁࢫ࣮ࣝࣉࢵࢺ࡞ 613V ᳨ฟἲࡢ⪃ࡀᚲせ࡛࠶ࡾࠊࡑࡢ᪉ἲㄽࢆᶍ⣴ࡋ࡚ ࠸ࡓࠋḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮1*6ࡢฟ⌧ࡣࠊᕧࣛ ࣈ࣮ࣛࣜࡢࢫࢡ࣮ࣜࢽࣥࢢࡣ᭱㐺ࡢ᪉ἲ࡛࠶ࡾࠊ ᮏᨭࡼࡾࡑࡢྍ⬟ᛶࢆ᳨ドࡍࡿࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 7,//,1*'1$ ࣛࣈ࣮ࣛࣜࡣ⣙ ಶయࡽࡢࢤ ࣀ࣒ '1$ ࡼࡾᵓᡂࡉࢀ࡚࠸ࡿࠋྛಶయ⏤᮶ '1$ ࡢ ୍㒊ࢆ 1*6 ࡼࡾゎᯒࡋࠊ┠ⓗ㑇ఏᏊᑟධࡉࢀ࡚ ࠸ࡿኚ␗ࢆࣁࢫ࣮ࣝࣉࢵࢺ᳨ฟࡍࡿࣉࣟࢪ࢙ࢡ ࢺࡢᨭࢆ⏦ㄳࡋࡓࠋᐇ㝿ࡣࠊࣛࣈ࣮ࣛࣜࡢ ༙ᩘࢧࣥࣉࣝࡘ࠸࡚ࠊ 㑇ఏᏊࡢࢫࢡ࣮ࣜࢽࣥࢢ ࢆ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 1*6 ゎᯒࡼࡿ 613V ᳨ฟ࠾࠸࡚ࠊ㝧ᛶഇ㝧ᛶ ࢆ༊ูࡍࡿᚲせ࡞ࠕ㝧ᛶࢡ࣮ࣟࣥࡢྜࠖཬࡧ ࠕࡑࢀࡽࢆ᳨ฟࡍࡿᚲせ࡞࣮ࣜࢻᩘࠖࢆ๓ࡢࣃ ࣟࢵࢺᐇ㦂᳨࡛ウࡋࡓࡇࢁࠊ ࢡ࣮ࣟࣥ㸯ಶ ࡢ㝧ᛶࢡ࣮ࣟࣥࡀᏑᅾࡍࢀࡤࠊ㏻ᖖࡢࢫࢡ࣮ࣜࢽࣥ ࢢ᳨࡛ฟྍ⬟࡛࠶ࡿࡀุ᫂ࡋࡓࠋࡑࡇ࡛ࠊࢤࣀ࣒ '1$ ࣛࣈ࣮ࣛࣜࡢ༙ᩘࢆ ಶయศ࡙ࡘ PL[ ࡋࠊࡑ ࡢྛࠎࢆࣉ࣐࣮ࣛタᐃࡋ࡚࠸ࡿࣂ࣮ࢥ࣮ࢻ࡛ศ ูࡍࡿࡼࡾయࡢࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜࠺ࡋ ࡓࠋ ᐇ㝿ࡣࠊᇶ♏◊✲ࡽᛂ⏝◊✲ࡲ࡛ᖜᗈ࠸ศ㔝 ࡛ゎᯒ࣭⏝ࡀᮇᚅࡉࢀࡿ㔜せ㑇ఏᏊ ಶࢆ㑅ู ࡋࠊࡑࢀࡽ㑇ఏᏊࡽྜィ ࢚࢟ࢯࣥࡘ࠸࡚ࠊ 7,//,1* ࢤࣀ࣒ࣛࣈ࣮ࣛࣜ୰Ꮡᅾࡍࡿᑟධኚ␗ ࡢࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡗࡓࠋࡇࢀࡽᶆⓗ㑇ఏᏊ࢚࢟ ࢯࣥࡢ⃰⦰ࡣࠊྜᡂ 51$ ࣋ࢺ㸦$JLOHQW♫࡛ྜᡂ㸧 ࡼࡾ⾜࠸ࠊࢩ࣮ࢡ࢚ࣥࢩࣥࢢࢧࣥࣉࣝࡋࡓࠋ ᮍ ゎ ᯒ ࡢ 㑇 ఏ Ꮚ ࡶ ṧ ࡉ ࢀ ࡚ ࠸ ࡿ ࡀ ࠊ '1$3.FV &U\SWRFKURPH)UL]]OHG)UL]]OHG㸲✀㢮 ࡢ㌿ᅉᏊ➼ࠊከࡃࡢ㑇ఏᏊࡘ࠸࡚ࢼࣥࢭࣥࢫኚ ␗యࡢ᳨ฟᡂຌࡋࡓࠋᮏἲࡼࡾᚓࡽࢀࡿኚ␗ಶ యࡣࠊࢫࢡ࣮ࣜࢽࣥࢢࡋࡓᶆⓗ㓄ิ௨እࡶணᮇ ࡏࡠኚ␗ࡀᑟධࡉࢀ࡚࠾ࡾࠊࡇࢀ➼ࡣṇᖖಶయࣂ ࢵࢡࢡࣟࢫࡋࢡ࣮ࣜࣥࢵࣉࡍࡿᚲせࡀ࠶ࡿࠋࡇࢀ ➼ࡢኚ␗యࡘ࠸࡚ ୡ௦ࡢࣂࢵࢡࢡࣟࢫࢆ⾜ࡗࡓ ᚋࠊ⾲⌧ᆺゎᯒࢆ⾜ࡗ࡚࠸ࡿࠋ࣐࢘ࢫ࡛ࡣ⬇ᛶ⮴Ṛ ࠶ࡿ࠸ࡣዷ࡞ࡿࣔࣀࡢࠊ࣓ࢲ࡛࢝ࡣࢾࣝኚ␗࡛ ࡶ YLDEOH ࠶ࡿ࠸ࡣዷᛶࢆᣢࡘ➼ࠊணእࡢ⾲⌧ᆺࡀ ほᐹࡉࢀ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡾࠊᴟࡵ࡚㧗࠸ࣁࢫ࣮ࣝࣉࢵࢺࢆ ᐇ⌧࡛ࡁࡓࠋᮏᡂᯝࡼࡾࠊࡼᑠࡉ࡞㑇ఏᏊࢆ 㝖ࡁࠊᕼᮃࡍࡿ㑇ఏᏊࡢ࣓ࢲ࢝ኚ␗యࢆᚓࡿᡭἲࢆ ☜❧࡛ࡁࡓࠋ㏫㑇ఏᏛⓗᡭἲࡢ☜❧ࡣࠊ࣓ࢲ࢝ࡢࣔ ࢹࣝᐇ㦂ື≀ࡋ࡚ࡢ㔜せᛶࢆቑຍࡉࡏࡿࡢࡳṆ ࡲࡽࡎࠊࡢ့ஙᐇ㦂ື≀ࢆ⿵ࡍࡿ࣓ࢲ࢝ࡢ᪂ࡓ ࡞ᐇ㦂⣔ࡢ㛤Ⓨ㔜せ࡞ᇶ┙ࢆ࠼ࡿࡶࡢ࡛࠶ࡿ ⪃࠼ࡽࢀࡿࠋ ࡇ ࡇ ᩘ ᖺ ࡢ 㛫 Transcription Activator Like Effector Nuclease (TALEN) ࡸ Clustered Regularly Interspaced Short Palindromic Repeat)/Cas9 (CRISPR) ➼ࡢࢤࣀ࣒⦅㞟ᢏ⾡ࡀ㛤Ⓨ࣭ᐇ⏝ࡉࢀ࡚ࡁࡓࠋ ࡇࢀ➼ࡢᢏ⾡ࡣᶆⓗ㑇ఏᏊࡢ≉ᐃ㓄ิdeletion ࢆᑟධ࡛ࡁࡿࡽࠊࢼࣥࢭࣥࢫኚ␗యࢆᐜ᫆ ᚓࡿࡇࡀ࡛ࡁࡿ᭷⏝࡞ᢏ⾡࡛࠶ࡿࠋࡋࡋ࡞ࡀ ࡽࠊࡇࢀࡽࢤࣀ࣒⦅㞟ᢏ⾡࡛ࡣࠊᑠࡉࡃ࡚ࡶᩘሷ ᇶࡢdeletion ࡀධࡗ࡚ࡋࡲ࠸ࠊ㸯ሷᇶኚ␗ࡢⅬ✺ ↛ኚ␗యࢆసᡂࡍࡿࡣࠊຠ⋡ⓗ࡞ࣀࢵࢡࣥ➼ ࠸ࡃࡘࡢ㞀ቨࡀ᭷ࡿ⪃࠼ࡽࢀࡿࠋᡃࠎࡀᶞ❧ ࡋࡓ7,//,1*ἲࡣࠊࢼࣥࢭࣥࢫኚ␗ࡢࡳ࡛࡞ࡃ㸯ሷ ᇶ⨨ࡢⅬ✺↛ኚ␗యࢆྠᚓࡿࡀ࡛ࡁࡿⅬ ࡛ࠊࡇࢀ➼ࢤࣀ࣒⦅㞟ᢏ⾡ࢆ┦⿵࡛ࡁࡿࡶࡢ࡛࠶ ࡿࠋ≉ࠊ ࠕࢤࣀ࣒ᨭࠖࡢᨭࡼࡾ☜❧ࡋࡓࣁ ࢫ࣮ࣝࣉࢵࢺ࣭ࢫࢡ࣮ࣜࢽࣥࢢἲࢆ☜❧࡛ࡁࡓ ࡣᴟࡵ࡚ព⩏῝࠸⪃࠼࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ↓ࡋ - 442 - බເᨭㄢ㢟㸸2014 ᖺᗘࠊ2015 ᖺᗘ ࢫࢸࣟࢻ⭷ཷᐜయࡢᶵ⬟ゎᯒࢆ୰ᚰࡋࡓ༸ᡂ⇍࣭༸ㄏᑟᶵᵓࡢゎ᫂ ᇶ┙◊✲㸦㹁㸧 ᚨඖ ಇఙ࣭㟼ᒸᏛᏛ㝔⌮Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ༸ᡂ⇍༸ࡣཷ⢭ྍ⬟࡞༸Ꮚࡢᙧᡂࡢࡓࡵࡢ㔜 せ࡞ࢫࢸࢵࣉ࡛࠶ࡾࠊࢫࢸࣟࢻ࣍ࣝࣔࣥࡢෆࠊࣉ ࣟࢤࢫࢸࣟࣥ㢮ࡼࡾᘬࡁ㉳ࡇࡉࢀࠊ༠ㄪⓗ㐍⾜ ࡍࡿࠋࡋࡋࠊࡇࢀࡽࡢ⌧㇟ࡣูࠎࡢ⤒㊰ࡼࡾㄏ ᑟࡉࢀࡿ᥎ᐃࡉࢀ࡚࠸ࡿࠋ༸ᡂ⇍ಁ㐍ᅉᏊࡢⓎぢ ࢆ➃⥴ࡋ࡚༸ᡂ⇍ㄏᑟᶵᵓࡘ࠸࡚ࡣゎ᫂ࡀ㐍ࡵ ࡽࢀࡓࡀࠊ༸ㄏᑟᶵᵓࡘ࠸࡚ࡣ᫂࡞Ⅼࡀከࡃ ṧࡉࢀ࡚࠸ࡿࠋࡲࡓࠊྠ㐍⾜ࡍࡿࡇࢀࡽࡢㄏᑟ⤒ ㊰ࢆ༊ูࡍࡿᡭẁࡀ↓ࡗࡓࡓࡵゎᯒࡣᅔ㞴࡛࠶ࡗ ࡓࠋ⏦ㄳ⪅ࡽࡣࢮࣈࣛࣇࢵࢩࣗࣥࣅ࡛࣎༸ᡂ ⇍࣭༸ㄏᑟྍ⬟࡞⏕యࢵࢭἲࢆ㛤Ⓨࡋࡓࠋࡇ ࡢᡭἲࡼࡾ⏕యෆ࡛༸ᡂ⇍ࡢࡳࢆㄏᑟࡉࢀࡓ≧ែ ࡢ༸ࠊ༸ᡂ⇍࣭༸ࡢ୧᪉ࡀㄏᑟࡉࢀࡓ༸ࢆึࡵ࡚ ㄪ〇ࡍࡿࡇࡀ࡛ࡁࡿࡼ࠺࡞ࡗࡓࠋࡇࡢࡇࡼ ࡾ༸ㄏᑟ≉␗ⓗ࡞㑇ఏᏊⓎ⌧ኚࢆᤊ࠼ࡿࡇ ࡀྍ⬟࡞ࡗࡓࠋ⏦ㄳ⪅ࡽࡣ᪤࣐ࢡࣟࣞࢆ ⏝࠸ࡓ⥙⨶ⓗ࡞㑇ఏᏊⓎ⌧ゎᯒࢆ㐍ࡵ࡚ࡁࡓࡀࠊᚓ ࡽࢀࡓࢹ࣮ࢱࡣ⌧ᛶࠊⓎ⌧ኚື⋡ၥ㢟ࡀឤࡌ ࡽࢀࡓࠋྠᮇࢤࣀ࣒ࢩࢡ࢚ࣥࢧ࣮ࢆ⏝࠸ࡓ RNA-seq ࡼࡿⓎ⌧ゎᯒᢏ⾡ࡀ☜❧ࡉࢀࠊᐇ⏝ࡢ ẁ㝵ධࡗࡓࠋᮏᨭ᥇ᢥࡉࢀࡓࡇࡽ࣐ࢡ ࣟࣞゎᯒࡢࢹ࣮ࢱ RNA-seq ゎᯒࡢࢹ࣮ࢱࢆ ẚ㍑ࡋࠊ⤫ྜࡍࡿࡇ࡛ࡼࡾṇ☜࡞Ⓨ⌧ኚືࢆᤊ࠼ ࡼ࠺⪃࠼ࡓࠋ 㸺ᨭࡢෆᐜ㸼 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᚓࡽࢀࡓ RNA-seq ゎᯒࡢࢹ࣮ࢱࡣྠ୍ࢭࢵࢺ ෆࡢ 4 ࠶ࡿ࠸ࡣ 6 ヨᩱ㛫࡛ࡢࢹ࣮ࢱࡢ୍⮴⋡ࡣᴟ ࡵ࡚㧗ࡃゎᯒࡀ㠀ᖖṇ☜⾜ࢃࢀࡓุᐃࡉࢀ ࡓࠋࡲࡓࠊⓎ⌧ኚື⋡ࡘ࠸࡚ࡶᴟ➃㧗࠸್ࢆ ♧ࡋࡓ㑇ఏᏊࡶ↓ࡃࠊࡇࡢⅬ࠾࠸࡚ࡶಙ㢗ᛶࡢ 㧗࠸ࢹ࣮ࢱࡀᚓࡽࢀࡓࡶࡢ᥎ᐃࡉࢀࡓࠋ୍᪉࡛ ࣐ࢡࣟࣞゎᯒࡢࢹ࣮ࢱ↷ྜࡋࡓ⤖ᯝࠊព እࡶࢇ୍⮴ࡣぢࡽࢀ࡞ࡗࡓࠋࡍ࡞ࢃࡕ ࣐ࢡࣟࣞゎᯒ࡛㑅ᢥࡋࡓ༸క࡞࠸Ⓨ⌧ ୖ᪼ࡍࡿ㑇ఏᏊ⩌ࡣ RNA-seq ゎᯒ࡛ࡣᮇᚅࡉࢀ ࡓⓎ⌧ୖ᪼ࡣࡳࡽࢀ࡞ࡗࡓࠋࡇࡢ⤖ᯝࢆ㋃ࡲ࠼ ࡚⮬↛᮲௳ୗ࡛ࡢヨᩱࢆຍ࠼ࡓ 6 ࢧࣥࣉ࡛ࣝࡢ ゎᯒࢆ౫㢗ࡋࡓࠋࡇࡢゎᯒ⤖ᯝࡶࢭࢵࢺෆ࡛ࡢࢹ ࣮ࢱࡢᏳᐃᛶࡣ༑ศ࡛࠶ࡗࡓࡀࠊࢭࢵࢺ㛫࡛ࡢࢹ ࣮ࢱࡢ୍⮴ࡀ᫂☜࡛ࡣ࡞ࡗࡓࠋࡑࡇ࡛ 2 ᖺ┠ ࡞ࡿ 2015 ᖺࡉࡽ㸰ࢭࢵࢺࡢゎᯒࢆ౫㢗ࡋࠊ ィ㸲ࢭࢵࢺࡢࢹ࣮ࢱ࡛ẚ㍑ࡍࡿࡇ࡞ࡗࡓࠋ ௨ୖࡢࡇࡽࠊᑡ࡞ࡃࡶᡃࠎࡢゎᯒ࠾࠸࡚ ࡣࢡࣟࣞゎᯒ࡛ᚓࡽࢀࡓࢹ࣮ࢱ RNA-seq ゎᯒࡼࡿࢹ࣮ࢱࡣࡁ࡞㝸ࡓࡾࡀ ࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࡢࡇࡽ୧ࢹ࣮ ࢱࢆ⤫ྜࡋ࡚Ⓨ⾲ࡍࡿࡇࡣᅔ㞴࡛࠶ࡿࡢ⤖ㄽ ⮳ࡗࡓࠋࢡࣟࣞゎᯒࡼࡿೃ⿵㑇ఏᏊࡢ 㑅ᢥࡘ࠸࡚ࡣᢞ✏‽ഛ୰࡛࠶ࡿࠋ⌧ᅾࠊ CRISR/Cas9 ࢩࢫࢸ࣒ࡼࡿೃ⿵㑇ఏᏊࡢ㑇ఏᏊ◚ ቯࡼࡿ㑇ఏᏊᶵ⬟ࡢド᫂ἲࢆᑟධ୰࡛࠶ࡿࡢ࡛ ༸ࡢ 3 ✀㢮ࡢ≧ែࠊࡍ࡞ࢃࡕ༸ᡂ⇍ࡶ༸ࡶㄏ ㉳ࡉࢀ࡚࠸࡞࠸≧ែ㸦ᮍฎ⌮㸧 ࠊ༸ᡂ⇍ࡢࡳㄏᑟࡉ ࢀ࡚࠸ࡿ≧ែ㸦ࢪ࢚ࢳࣝࢫࢳࣝ࣋ࢫࢺ࣮ࣟࣝࠊࢸ ࢫࢺࢫࢸࣟࣥฎ⌮㸧 ࠊ༸ᡂ⇍࣭༸ࡢ୧᪉ࡀㄏᑟࡉ ࢀ࡚࠸ࡿ≧ែ㸦㨶㢮ࡢኳ↛ࡢ༸ᡂ⇍ㄏ㉳࣍ࣝࣔࣥ ࡛࠶ࡿ 17,20ș-DHP ฎ⌮㸧ࡢィ 4 ✀㢮ࡢ༸ᕢࡽ ᚓࡽࢀࡓ RNA ヨᩱࢆ 1 ࢭࢵࢺࡋ࡚ RNA-seq ゎ ᯒࢆ౫㢗ࡋࡓࠋ RNA-seq ゎᯒ࡛ᚓࡽࢀࡓೃ⿵㑇ఏᏊ⩌ࡘ࠸࡚ ࡣᶵ⬟ド᫂ࡲ࡛ྵࡵࡓෆᐜࡋ࡚ᩘᖺᚋࡢⓎ⾲ࢆ ┠ᣦࡋࡓ࠸ࠋ RNA ヨᩱ㛫࡛ࡢẚ㍑ࢆ⾜࠺ࡓࡵ 2014, 2015 ᖺᗘ ࡘ࠸࡚ྛ 2 ࢭࢵࢺࠊィ㸲ࢭࢵࢺࡢࢩ࣮ࢡ࢚ࣥࢫゎᯒ ࢆᐇࡋ࡚࠸ࡓࡔ࠸ࡓࠋ2 ࢭࢵࢺ┠௨㝆ࡣ⮬↛᮲௳ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣐ࢡࣟࣞゎᯒࡣ␗㉁ࡢࢹ࣮ࢱࡀᚓࡽࢀࡓ ࡇ࡛᪂ࡓ࡞ೃ⿵㑇ఏᏊ⩌ࡢࣜࢫࢺࡀᚓࡽࢀࡓࠋ ᚋࠊq-PCR ࡛ࡢⓎ⌧ࣞ࣋ࣝࡢ᳨ドࢆ⾜ࡗ࡚࠸ࡁࠊ ࢡࣟࣞ RNA-seq ࡢࢹ࣮ࢱࢆẚ㍑ࡋࠊೃ⿵㑇 ఏᏊࡢ㑅ᢥࢆ㐍ࡵ࡚࠸ࡃࠋ ୗ࡛ࡢⓎ⌧ኚࢆ☜ㄆࡍࡿࡓࡵࠊ࣍ࣝࣔࣥฎ⌮ࢆ ⾜ࢃ࡞࠸⮬↛㓄᮲௳ୗ࡛ࡢࢧࣥࣉࣜࣥࢢヨᩱࡶ ຍ࠼ࡓࠋࢮࣈࣛࣇࢵࢩࣗࡣ᪥ࡢฟࡶ⏘༸ࢆ 㛤ጞࡍࡿࡢ࡛㣫⫱᮲௳ୗ࡛ࡢⅬⅉᩘ㛫๓Ⅼⅉ┤ ๓ࡢ༸ᕢࢆ༸ᡂ⇍ᮇࠊ༸ᮇࡢヨᩱࡋ࡚᥇ྲྀࡋࠊ RNA ࢆᢳฟࡋࡓࠋࡇࢀࡽ 2 ࢧࣥࣉࣝࢆຍ࠼ࡓィ 6 ࢧ ࣥࣉࣝࡋࡓࠋ ⌧ᅾࠊ㸲ࢭࢵࢺ┠ࡢゎᯒࡀ㐍⾜୰࡛࠶ࡾࠊࢹ࣮ࢱ ࡀᚓࡽࢀḟ➨ࠊ㸲ࢭࢵࢺయ࡛ࡢⓎ⌧ኚືゎᯒࢆ⾜ ࠸ࠊRNA-seq ゎᯒ࠾ࡅࡿ༸ㄏᑟ㑇ఏᏊࡢ᭱⤊ ⓗ࡞㑅ᢥࢆ⾜࠸ࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᇳ➹୰ - 443 - බເᨭㄢ㢟㸸2010 ᖺᗘ ࣃ࣮࢟ࣥࢯࣥ࠾ࡼࡧㄆ▱ᶵ⬟㛵㐃ศᏊࣃ࣮ࢯࢼࣝࢤࣀ࣒ゎᯒ ᪂Ꮫ⾡㡿ᇦ◊✲ ᡞ⏣ 㐩ྐ࣭⚄ᡞᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ࣃ࣮࢟ࣥࢯࣥ(PD)ࡣࠊ୰㧗ᖺⓎࡋࠊࢻࣃ࣑ࣥ ࢽ࣮ࣗࣟࣥࡢኚᛶࡼࡾᡓࠊ➽ᅛ⦰࡞㐠ື㞀ᐖ ࢆ≧ࡍࡿ㢖ᗘࡢ㧗࠸⚄⤒ᝈ࡛࠶ࡾࠊ㒊ศ ࢆ༨ࡵࡿᘼⓎᛶ PD ࡛ࡣᝈឤཷᛶ㑇ఏᏊࡣ᫂ࡽ ࡛࡞࠸ࠋࠕㄆ▱ᶵ⬟㑇ఏᏊ㺃ศᏊࠖࡣ㸰㸯ୡ⣖ࡢ⚄ ⤒⛉Ꮫࡢ୰ᚰⓗࢸ࣮࣐ࡢ㸯ࡘ࡛࠶ࡾࠊ⯡ࡢ▱ⓗ⬟ ຊ㛵ࢃࡿ୍⯡▱⬟ᅉᏊ g ࡣࠊᮏ㉁ⓗከᅉᏊ㑇ఏ ⓗ࡛࠶ࡿࠋࡲࡓ IQ ࡀ 70 ᮍ‶࡛࠶ࡿ⢭⚄㐜ࡣࠊ୍ ⯡ேཱྀࡢ 3㸣⛬ᗘࡢ㢖ᗘࡉࢀࠊᴟࡵ࡚㢖ᗘࡢ㧗࠸ ᝈ⩌࡛࠶ࡿࠋᮏ◊✲࡛ࡣࠊ௦⾲ⓗ࡞⚄⤒㞴ࡢࣃ࣮ ࢟ࣥࢯࣥ࠾ࡼࡧ୍⯡▱⬟ᅉᏊࡸ⢭⚄㐜࡞ࡢㄆ ▱ᶵ⬟㛵㐃ศᏊ↔Ⅼࢆ࠾ࡁࠊࣃ࣮࢟ࣥࢯࣥࠊ⢭ ⚄㐜㺃⮬㛢ࡢែゎ᫂ࠊ⒪ἲࡢ㛤Ⓨࠊ▱⬟Ⓨ⌧ ࡢศᏊ࣓࢝ࢽࢬ࣒ࡢゎ᫂ࢆ┠ⓗࡍࡿࠋᮏィ⏬࡛ࡣ ≉ࠊᙉ࠸ࡶࡋࡃࡣከᩘࡢ㑇ఏせᅉࡀ㞟✚ࡋ࡚࠸ࡿ ணࡉࢀࡿᐙ᪘㞟✚ᝈ⪅ࡢ࢚ࢡࢯ࣮࣒ࢩ࣮ࢡ࢚ࣥ ࢫࢆ࠾ࡇ࡞࠸ࠊࣃ࣮࢟ࣥࢯࣥ㑇ఏᏊࢆྠᐃࡍࡿࡇ ࢆ┠ᣦࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᐙ᪘ᛶࣃ࣮࢟ࣥࢯࣥᝈ⪅ 6 ᳨యࡢ࢚ࢡࢯ࣮࣒ࢩ࣮ ࢡ࢚ࣥࢫᐇ㦂ゎᯒࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 6 ᳨యࡢ coverage ࡣࠊx423, x204, x461, x224, x200, x220 ࡛࠶ࡗࡓࠋMapping rate ࡣࠊ86%, 86%, 86%, 85%, 84%, 84%࡛࠶ࡗࡓࠋPCR duplicate ࡣࠊ 10.5%, 5.6%, 12.8%, 7,1%, 7.5%, 8.4%࡛࠶ࡗࡓࠋ Insert size ࡢศᕸࡣࠊ213(SD 64), 230 (67), 205 (60), 229 (70), 209 (63), 213 (65)࡛࠶ࡗࡓࠋDB Ⓩ㘓ࡉ ࢀ࡚࠸࡞࠸ rare SNV ᩘࡣࠊ37918, 23455, 57363, 25195, 32372, 40746 ࡛࠶ࡗࡓࠋIndel ᩘࡋ࡚ࡣࠊ rare coding ࢇࡋ࡚ࡣࠊ71, 45, 57, 51, 50, 51 ࡛ ࠶ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ୖグࡢࢹ࣮ࢱࡢඹ㏻ SNV ࡞᳨ウࡋࡓࡀࠊࡇࢀࡽ ࡢࡳ࡛ࡣࣃ࣮࢟ࣥࢯࣥ㑇ఏᏊྠᐃ⮳ࡽ࡞ࡗࡓࠋ ᚋࠊูᐇ㦂῭ࡳࡢࣃ࣮࢟ࣥࢯ࢚ࣥࢡࢯ࣮࣒ゎ ᯒᩘⓒ᳨యࡢࢹ࣮ࢱࢆཧ↷ࡋࠊࡈᨭࡢࢹ࣮ࢱࢆྜ ࢃࡏ࡚ゎᯒࡍࡿࡇࡼࡾࠊ᪂つࡢࣃ࣮࢟ࣥࢯࣥ 㑇ఏᏊࢆⓎぢ࡛ࡁࡿྍ⬟ᛶࡀ࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 444 - බເᨭㄢ㢟㸸2011 ᖺᗘ ࣃ࣮࢟ࣥࢯࣥ࠾ࡼࡧㄆ▱ᶵ⬟㛵㐃ศᏊࣃ࣮ࢯࢼࣝࢤࣀ࣒ゎᯒ ᪂Ꮫ⾡㡿ᇦ◊✲ ᡞ⏣ 㐩ྐ࣭⚄ᡞᏛᏛ㝔་Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ᯒࡢࢤࣀ࣒་Ꮫᨭ ࣃ࣮࢟ࣥࢯࣥ(PD)ࡣࠊ୰㧗ᖺⓎࡋࠊࢻࣃ࣑ ࣥࢽ࣮ࣗࣟࣥࡢኚᛶࡼࡾᡓࠊ➽ᅛ⦰࡞㐠ື㞀 ᐖࢆ≧ࡍࡿ㢖ᗘࡢ㧗࠸⚄⤒ᝈ࡛࠶ࡾࠊ㒊 ศࢆ༨ࡵࡿᘼⓎᛶ PD ࡛ࡣᝈឤཷᛶ㑇ఏᏊࡣ᫂ࡽ ࡛࡞࠸ࠋ୍᪉ࠕㄆ▱ᶵ⬟㑇ఏᏊ㺃ศᏊࠖࡣ㸰㸯ୡ ⣖ࡢ⚄⤒⛉Ꮫࡢ୰ᚰⓗࢸ࣮࣐ࡢ㸯ࡘ࡛࠶ࡾࠊ⯡ࡢ ▱ⓗ⬟ຊ㛵ࢃࡿ୍⯡▱⬟ᅉᏊ g ࡣࠊᮏ㉁ⓗከᅉ Ꮚ㑇ఏⓗ࡛࠶ࡿࠋࡲࡓ IQ ࡀ 70 ᮍ‶࡛࠶ࡿ⢭⚄㐜 ࡣࠊ୍⯡ேཱྀࡢ 3㸣⛬ᗘࡢ㢖ᗘࡉࢀࠊᴟࡵ࡚㢖ᗘࡢ 㧗࠸ᝈ⩌࡛࠶ࡿࠋᮏ◊✲࡛ࡣࠊ௦⾲ⓗ࡞⚄⤒㞴 ࡢࣃ࣮࢟ࣥࢯࣥ࠾ࡼࡧ୍⯡▱⬟ᅉᏊࡸ⢭⚄㐜࡞ ࡢㄆ▱ᶵ⬟㛵㐃ศᏊ↔Ⅼࢆ࠾ࡁࠊࣃ࣮࢟ࣥࢯࣥ ࠊ⢭⚄㐜㺃⮬㛢ࡢែゎ᫂ࠊ⒪ἲࡢ㛤Ⓨࠊ▱ ⬟Ⓨ⌧ࡢศᏊ࣓࢝ࢽࢬ࣒ࡢゎ᫂ࢆ┠ⓗࡍࡿࠋᮏィ ⏬࡛ࡣ≉ࠊㄆ▱ᶵ⬟㛵㐃ࡍࡿ㑇ఏᏊࢆྠᐃࡍࡿ ࡇࢆ┠ᣦࡋࠊIQ ࡢᕪࡀ㢧ⴭ࡞୍⮴୍༸ᛶ⏕ඣࠊ ࠾ࡼࡧ⢭⚄㐜ᐙ⣔ࢆ⏝࠸ࡓつᶍࢤࣀ࣒ࢩ࣮ࢣ ࣥࢩࣥࢢࢆ┠ⓗࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 IQ ᕪࡢ᭱ࡶࡁ࠸୍⮴୍༸ᛶ⏕ඣ 2 ⤌ 4 ᳨య ࡢࢤࣀ࣒㸦IQ ᕪࡣࠊ1 ⤌ࡣ 45ࠊࡶ࠺ 1 ⤌ࡣ 30㸦1 SD = 15㸧㸧ࠊ⢭⚄㐜ᐙ⣔ 2 ᐙ⣔ 6 ᳨యࡢ࢚ࢡࢯ࣮࣒ ࡢࢩ࣮ࢣࣥࢩࣥࢢ㸦1 ᐙ⣔ࡣẕぶᘵࡀࠊࡶ࠺ 1 ᐙ⣔ࡣᘵࡀ⨯ᝈࡋ࡚࠸ࡿᐙ⣔㸧ࠊ࠾ࡼࡧࡑࡢሗゎ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⢭⚄㐜ᐙ⣔ࡢ࢚ࢡࢯ࣮࣒ࢩ࣮ࢣࣥࢩࣥࢢࡼ ࡾࠊ1 ᐙ⣔ࡽ᪤▱ࡢ⢭⚄㐜ཎᅉ㑇ఏᏊୖࡢኚ␗ࢆ ぢฟࡋࡓࠋࡶ࠺ 1 ᐙ⣔ࡽࡣࠊ᪂つࡢཎᅉ㑇ఏᏊೃ ⿵ࢆྠᐃࡋࡓࠋ୍⮴୍༸ᛶ⏕ඣࡢࢤࣀ࣒ࢩ ࣮ࢣࣥࢩࣥࢢࡣࠊヨᩱᥦ౪ඖ࡛ࡢ⌮ጤဨ࡛ᢎㄆ ࡀᚓࡽࢀࡎࠊᨭࢆ᩿ᛕࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ཎᅉ᫂ࡢ⢭⚄㐜㸯ᐙ⣔ࡘ࠸࡚ࡢཎᅉ㑇ఏᏊ ࡢྠᐃᡂຌࡋࠊ☜ᐃデ᩿ࡍࡿࡇࡀ࡛ࡁࡓࠋࡶ࠺ 㸯ࡘࡢཎᅉ᫂ࡢ⢭⚄㐜ᐙ⣔ࡽྠᐃࡉࢀࡓ⢭⚄ 㐜ཎᅉ㑇ఏᏊೃ⿵ࡘ࠸࡚ࡣࠊࡇࢀࡲ࡛ࡢ⢭⚄ 㐜ᝈ⪅ࡢኚ␗᳨⣴ࢆ⾜ࡗ࡚࠸ࡿࡀࠊࡢࡇࢁⓎ ぢࡣ⮳ࡗ࡚࠸࡞࠸ࠋࡉࡽ࡞ࡿゎᯒࢆ⾜࠺ࡇࡼ ࡾࠊ᪂つࡢ⢭⚄㐜ཎᅉ㑇ఏᏊࡢྠᐃࡘ࡞ࡀࡿࡇ ࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 445 - බເᨭㄢ㢟㸸2014 ᖺᗘ ຍ㱋⬻࠾ࡅࡿ࣓ࢰᅇ㊰ࡢᶵ⬟㞀ᐖ ᪂Ꮫ⾡㡿ᇦ◊✲ Ẋᇛ ள▮Ꮚ࣭༓ⴥᏛᏛ㝔⸆Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 Ꮫ⩦ࡼࡗ࡚ᙧᡂࡉࢀࡿグ᠈ࡣࠊᏳᐃ࡞▷ᮇグ᠈ ࡽᏳᐃ࡞㛗ᮇグ᠈ᅛᐃࡉࢀࡿࠋ㛗ᮇグ᠈ࡢ ᙧᡂࡣࠊ≉ᐃࡢ㑇ఏᏊⓎ⌧࠾ࡼࡧࢱࣥࣃࢡ㉁ྜᡂ ࡀᚲせ࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋࡇࡢࡼ࠺࡞Ᏻᐃ ࡞グ᠈ࡢᙧᡂ⬟ࡣຍ㱋కࡗ࡚పୗࡍࡿࡀࠊࡑࡢศ Ꮚ࣓࢝ࢽࢬ࣒ࡣࡲࡔゎ᫂ࡉࢀ࡚࠸࡞࠸ࠋᡃࠎࡣࢩࣙ ࢘ࢪࣙ࢘ࣂ࢚ࡢႥぬグ᠈ࢩࢫࢸ࣒ࢆࣔࢹࣝ⣔ࡋ࡚ ຍ㱋౫Ꮡⓗ࡞グ᠈ᙧᡂ⬟ࡢኚࢆゎᯒࡋࠊࢱࣥࣃࢡ ㉁ྜᡂ౫Ꮡⓗ࡞㛗ᮇグ᠈ࡢᙧᡂࡀຍ㱋క࠸పୗࡋ ࡚࠸ࡿࡇࢆ᫂ࡽࡋ࡚ࡁࡓࠋ୍᪉࡛ࠊ▷ᮇグ᠈ ࡢᙧᡂࡸࢱࣥࣃࢡ㉁ྜᡂ㠀౫Ꮡⓗ࡞㛗ᮇグ᠈ࡢᙧᡂ ࡣࠊຍ㱋క࠸㢧ⴭ࡞ኚࡀぢࡽࢀ࡞ࡗࡓࠋࡇࡢ ࡇࡼࡾࠊࢱࣥࣃࢡ㉁ྜᡂ౫Ꮡⓗ࡞㛗ᮇグ᠈ࡢᙧᡂ 㐣⛬ࡀຍ㱋ࡼࡾ≉␗ⓗ㞀ᐖࢆཷࡅ࡚࠸ࡿࡇࡀ ♧၀ࡉࢀࡓTonoki and Davis.,JNeurosci.,2015ࠋࡑࡇ ࡛ࠊ⬻ෆࡢ㑇ఏᏊⓎ⌧ࡸ㛗ᮇグ᠈ࡢᙧᡂ㐣⛬࡛Ⓨ⌧ ኚࡍࡿ㑇ఏᏊࣉࣟࣇࣜࣥࢢࢆⱝ㱋ಶయ⪁㱋 ಶయ࡛ẚ㍑ゎᯒࢆ⾜࠸ࠊೃ⿵㑇ఏᏊ⩌ࡽ㑇ఏᏛⓗ ࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜࠺ࡇ࡛ຍ㱋క࠺グ᠈పୗࡢ ཎᅉ࡞ࡿ㈐௵㑇ఏᏊ⩌ࢆ⥙⨶ⓗྠᐃࡍࡿࡇࢆ ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 㸦㸯㸧ຍ㱋క࠺㑇ఏᏊࣉࣟࣇࣜࣥࢢࡢኚ㸸 ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ⱝ㱋ಶయ㸦 ᪥㱋㸧⪁㱋ಶయ㸦 ᪥㱋㸧ࡢ⬻ 51$ ࢆࡑࢀࡒࢀ ࢧࣥࣉࣝࡎࡘᅇࡋࠊ 51$TXDOLW\FKHFN ࡢᚋࠊḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮ゎᯒ 51$VHTࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸦㸰㸧㛗ᮇグ᠈ᙧᡂ㐣⛬࡛ኚࡍࡿ㑇ఏᏊࣉࣟࣇ ࣜࣥࢢ㸸㛗ᮇグ᠈ᙧᡂࡣࠊ⧞ࡾ㏉ࡋᏛ⩦ࡀᚲせ ࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋⱝ㱋ಶయ㸦 ᪥㱋㸧ࠊ⪁ 㱋ಶయ㸦 ᪥㱋㸧ࡑࢀࡒࢀ࠾࠸࡚ࠊ⧞ࡾ㏉ࡋᏛ⩦ ᚋࠊ 㛫࣭ 㛫ࡢ⬻ 51$ ࢆࡑࢀࡒࢀ ࢧࣥࣉࣝ ࡎࡘᅇࡋࠊ51$TXDOLW\FKHFN ࡢᚋࠊḟୡ௦ࢩ࣮ࢡ ࢚ࣥࢧ࣮ゎᯒ51$VHTࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 㸦㸯㸧ຍ㱋క࠺㑇ఏᏊࣉࣟࣇࣜࣥࢢኚࡢゎ ᯒࡼࡾࠊ⪁ࡼࡾ⬻ෆ࡛Ⓨ⌧ࡀୖ᪼ࡍࡿ㑇ఏᏊ ⩌㸦ORJ)ROG&KDQJH)&㸧JHQHVࠊ ಸ௨ୖୖ᪼ ࡍࡿ㑇ఏᏊ⩌㸦ORJ)&㸧JHQHVࠊ⪁ࡼࡾ⬻ ෆ࡛Ⓨ⌧ࡀపୗࡍࡿ㑇ఏᏊ⩌ORJ)&!JHQHVࠊ ಸ௨ୖపୗࡍࡿ㑇ఏᏊ⩌ORJ)&!JHQHV ࡢࣜࢫ ࢺࢆᚓࡓࠋ 㸦㸰㸧㛗ᮇグ᠈ᙧᡂ㐣⛬࡛ኚࡍࡿ㑇ఏᏊࣉࣟࣇ ࣜࣥࢢࡢኚゎᯒࡼࡾࠊ⧞ࡾ㏉ࡋᏛ⩦ᚋ 㛫 Ⓨ⌧㔞ࡀୖ᪼ࡍࡿ㑇ఏᏊ⩌ࢆⱝ㱋 JHQHVࠊ⪁㱋 JHQHVࠊపୗࡍࡿ㑇ఏᏊ⩌ࢆⱝ㱋 JHQHVࠊ⪁㱋 JHQHV ࡢࣜࢫࢺࢆᚓࡓࠋࡇࢀࡽ㑇ఏᏊ⩌ࡘ࠸࡚ *HQH2QWRORJ\ ゎ ᯒ ࢆ ⾜ ࡗ ࡓ ⤖ ᯝ ࠊ ͇PLFURWXEXOH ELQGLQJ͇͇SURWHLQ VHULQHWKUHRQLQH NLQDVH DFWLYLW\͇͆FDOPRGXOLQELQGLQJ͇*27HUP ࠾࠸ ࡚ඃ㧗࠸ฟ⌧㢖ᗘࢆᣢࡘࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ୍᪉࡛ྠᵝࡢ *27HUP ࡣ⪁㱋ಶయ࠾࠸࡚ࠊࡑࡢⓎ⌧ 㢖ᗘࡣⱝ㱋ಶయẚ㍑ࡋ࡚ඃప࠸ࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡇࢀࡽࡢࡇࡽࠊ⪁㱋ಶయ࠾࠸࡚ࡣ ඛิᣲࡋࡓ *27HUP ྵࡲࢀࡿ㑇ఏᏊ⩌ࡀኚࡋ࡞ ࠸ࡇࡀࠊࢱࣥࣃࢡ㉁ྜᡂ౫Ꮡⓗ࡞㛗ᮇグ᠈ࡀపୗ ࡍࡿཎᅉࡢ୍ࡘ࡛࠶ࡿࡇࡀ♧၀ࡉࢀࡓࠋ ࡇࢀࡽࡢゎᯒࢆඖ⌧ᅾࠊೃ⿵㑇ఏᏊ⩌ࡢ⥙⨶ⓗ 㑇ఏᏛࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡗ࡚࠸ࡿࠋලయⓗࡣࠊ ຍ㱋క࠸ኚࡍࡿ㑇ఏᏊ⩌ࡇࢀࡲ࡛グ᠈ᙧᡂ 㛵ࡍࡿ⥙⨶ⓗ㑇ఏᏛⓗࢫࢡ࣮ࣜࢽࣥࢢࡼࡾᚓࡽ ࢀࡓ㑇ఏᏊ⩌㸦:DONLQVKDZHWDO*HQHWLFV㸧 ࢆẚ㍑ゎᯒࡋࡓࠋࡇࢀࡼࡾࠊDຍ㱋ࡼࡾ㑇ఏᏊⓎ ⌧㔞ࡀୖ᪼ࡍࡿࠊࡘグ᠈పୗ㛵ࡍࡿྍ⬟ᛶࡀ ࠶ࡿ㑇ఏᏊ⩌ࢆ 㑇ఏᏊ㸦*HQHQDPH 6$ DPRQ J\FEHWD%FS㸧ࠊEຍ㱋ࡼࡾ㑇ఏᏊⓎ⌧㔞 ࡀపୗࡍࡿࠊࡘグ᠈పୗ㛵ࡍࡿྍ⬟ᛶࡀ࠶ࡿ 㑇ఏᏊ⩌ 㑇ఏᏊ㸦*HQHQDPH JDODFWRNLQDVH NQRFNGRZQ JOXWDPDWHUHFHSWRUELQGLQJSURWHLQ &*&* ➼㸧ࢆྠᐃࡋࡓࠋ⌧ᅾࠊ㸦D㸧࡛ྠ ᐃࡉࢀࡓ㑇ఏᏊ⩌ࡘ࠸࡚ࠊLሙᡤ≉␗ⓗ㑇ఏᏊ ࢆⓎ⌧ࡉࡏࡿࡇࡀྍ⬟࡞ *DO8$6 㑇ఏᏊⓎ⌧⣔࠾ ࡼࡧLLᮇ≉␗ⓗ㑇ఏᏊࢆⓎ⌧ࡉࡏࡿࡇࡀ ྍ⬟࡞ *DO8$6 㑇ఏᏊⓎ⌧⣔࡛࠶ࡿ͆JHQHVZLWFK ࢩࢫࢸ࣒͇㸦5RPDQHWDO31$6㸧ࢆ⏝࠸࡚ࠊ ⚄⤒⣽⬊≉␗ⓗࡘᮇ≉␗ⓗࡑࢀࡒࢀࡢ㑇ఏᏊ ࢆࣀࢵࢡࢲ࢘ࣥࡉࡏࡓಶయࢆసᡂࡋࠊࡉࡽࡑࢀࡒ ࢀࡢಶయ࠾ࡅࡿグ᠈⬟ࡢ ᐃࢆ㐍ࡵ࡚࠸ࡿࠋࡇࢀ ࡲ࡛ࠊDPRQ ࡘ࠸࡚ⱝ㱋ಶయ࣭⪁㱋ಶయ୧⪅࠾ ࠸࡚ࣀࢵࢡࢲ࢘ࣥࡉࡏࡓಶయ࡛グ᠈ࡢୖ᪼ࡀࡳࡽࢀ ࡓࡇࡽࠊຍ㱋ࡼࡾⓎ⌧㔞ࡀୖ᪼ࡍࡿࡇ࡛グ ᠈ࡢపୗᐤࡍࡿ㑇ఏᏊೃ⿵ࡢ୍ࡘ࡛࠶ࡿ⪃࠼ ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᨭࡢᡂᯝࡼࡗ࡚ࠊຍ㱋క࠸⬻ෆ࡛Ⓨ⌧పୗࡍ ࡿࡇ࡛グ᠈పୗࡢཎᅉ࡞ࡿ㑇ఏᏊ⩌ࠊຍ㱋క ࠸⬻ෆ࡛Ⓨ⌧ୖ᪼ࡍࡿࡇ࡛グ᠈పୗࡢཎᅉ࡞ࡿ 㑇ఏᏊ⩌ࠊ⧞ࡾ㏉ࡋᏛ⩦ᚋⓎ⌧ࢇࡍࡿ㑇ఏᏊ ⩌ࠊ⪁㱋ಶయ࠾࠸࡚⧞ࡾ㏉ࡋᏛ⩦ᚋⓎ⌧ኚࡋ ࡞࠸ࡇ࡛ࢱࣥࣃࢡ㉁ྜᡂ౫Ꮡⓗ࡞㛗ᮇグ᠈ᙧᡂప ୗࡢཎᅉ࡞ࡿ㑇ఏᏊ⩌ࢆᚓࡿࡇࡀ࡛ࡁࡓࠋୖグ ࡢࡼ࠺ࠊࡇࢀࡽࡢ㑇ఏᏊ⩌ࡽ㡰ḟ⥙⨶ⓗ㑇ఏᏛ ࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜࠺ணᐃ࡛࠶ࡿࠋࡇࢀࡽࡢ⤖ᯝ ࡽࠊ⪁ࡶ࡞࠺グ᠈పୗࡢཎᅉ࡞ࡿ㑇ఏᏊ⩌ ࢆຠ⋡ⓗྠᐃࡍࡿࡇࡀྍ⬟࡞ࡿ⪃࠼ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾࠊ‽ഛ୰ - 446 - බເᨭㄢ㢟㸸2015 ᖺᗘ ⪁క࠺௦ㅰᜏᖖᛶࡢ◚⥢ࡼࡿグ᠈పୗᶵᵓࡢゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ Ẋᇛ ள▮Ꮚ࣭༓ⴥᏛᏛ㝔⸆Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 Ꮫ⩦ࡸグ᠈ᶵ⬟ࡣࠊຍ㱋ࡶ࡞ࡗ࡚పୗࡍࡿࠋ㑇 ఏⓗࡶࡋࡃࡣ⎔ቃⓗᑑࢆ᧯సࡋࡓ㛗ᑑಶయࡣࠊ ຍ㱋ᛶグ᠈పୗࡢⓎࡀ㐜ࢀࡿࡇࡀ✀ࢆ㏻ࡌ࡚▱ ࡽࢀ࡚࠸ࡿࠋࡓ࠼ࡤࠊ࣮࢝ࣟࣜไ㝈ࢆ࠾ࡇ࡞ࡗࡓ ಶయࡸᡂ㛗࣍ࣝࣔࣥࢆపୗࡉࡏࡓಶయ࡛ࡣࠊ㛗ᑑ࡛ ࠶ࡿࡔࡅ࡛࡞ࡃ⪁ಶయ࠾ࡅࡿグ᠈పୗࡀᢚไࡉ ࢀࡿࠋᑑไᚚ㛵ࡍࡿ◊✲ࡣࠊࣥࢫࣜࣥࢩࢢࢼ ࣝ⤒㊰ࢆ୰ᚰ⾜ࢃࢀࠊࡇࢀࡽࡢᑑไᚚᶵᵓࡣ⥺ ࡽࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ࠊ࣐࢘ࢫ⮳ࡿ㐍ࡢ㐣⛬ ࡛ಖᏑࡉࢀ࡚࠸ࡿࠋ့ங㢮࠾࠸࡚ࡣຍ㱋ࡶ࡞ ࠸ࠊࣥࢫࣜࣥศἪ⬟ࡢపୗࡸࣥࢫࣜࣥᢠᛶࡢ ቑ࡞ࠊ௦ㅰᜏᖖᛶࡀ◚⥢ࡍࡿࡇࡶ▱ࡽࢀ࡚࠸ ࡿࠋࡇࢀࡽࡢ▱ぢࡼࡾࠊຍ㱋ࡶ࡞࠺⏕యෆࡢ௦ ㅰᜏᖖᛶࡢ◚⥢ࡀグ᠈ᶵᵓࢆኚࡉࡏࠊຍ㱋ᛶグ᠈ పୗࢆㄏ㉳ࡍࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿ ᡃࠎࡣࡇࢀࡲ࡛ࠊࢩࣙ࢘ࢪࣙ࢘ࣂ࢚Ⴅぬグ᠈ࢆ ࣔࢹࣝࡋ࡚ࠊ⪁ಶయ࡛ࡣᏳᐃ࡞グ᠈ࡀᙧᡂࡉࢀ ࡞࠸ࡇࠊグ᠈ᅛᐃࡢࣉࣟࢭࢫࢆᢸ࠺ᶵ⬟ࡀపୗࡋ ࡚࠸ࡿࡇࢆ᫂ࡽࡋ࡚ࡁࡓ㸦7RQRNL$DQG'DYLV 5/31$6㸧ࠋໝ࠸㟁Ẽ่⃭ࢆ⤌ࡳྜࢃࡏ ࡓ㐃ྜᏛ⩦ࡼࡾ⋓ᚓࡉࢀࡓグ᠈ࡣࠊ㛫ࡢ⤒㐣 ࡶᛀ༷ࡉࢀࡿࠋ⪁ಶయ࡛ࡣࠊグ᠈ࢆ⋓ᚓࡍࡿ ⬟ຊࡣⱝ㱋ಶయẚ㍑ࡋ࡚㢧ⴭపୗࡋ࡞࠸ࡀࠊグ ᠈ࢆ⥔ᣢ࣭ᅛᐃࡍࡿ⬟ຊࡀపୗࡋ࡚࠸ࡿࡇࡀ♧၀ ࡉࢀࡓࠋࡲࡓࠊᡃࠎࡣయෆࡢᜏᖖᛶグ᠈ᶵ⬟ࡢ㛵 ࡘ࠸࡚ࠊ௨ୗࢆ᫂ࡽࡋ࡚ࡁࡓࠋ㑇ఏᏛ ⓗࣥࢫࣜࣥ⏘⏕ࢆ୍㐣ⓗᢚไࡋࡓࣁ࢚ಶయ ࠾࠸࡚Ⴅぬグ᠈ࢆ ᐃࡋࡓࡇࢁࠊᏛ⩦ᶵ⬟ࡣṇᖖ ࡔࡀグ᠈ࢆ⥔ᣢࡍࡿ⬟ຊࡀపୗࡍࡿࠋࣥࢫࣜ ࣥ⏘⏕ࢆ⪁㱋ಶయ࡛ྠᵝᢚไࡉࡏ࡚ࡶࠊグ᠈పୗ ࡣஹ㐍ࡋ࡞࠸ࠋ㸦㸧ࣥࢫࣜࣥᵝ࣌ࣉࢳࢻࡢ୍㒊ࡸ ࡑࡢ㛵㐃ᅉᏊࡢⓎ⌧㔞ࡣࠊຍ㱋ࡼࡾపୗࡍࡿࠋ ࡇࢀࡲ࡛ࡢ⤖ᯝࢆ㋃ࡲ࠼ࠊᮏ◊✲ㄢ㢟࡛ࡣయෆࡢ ௦ㅰᜏᖖᛶࡢ◚⥢ࡀグ᠈ᶵ⬟࠼ࡿᙳ㡪ࢆ᫂ࡽ ࡍࡿࡇ࡛ࠊຍ㱋ᛶグ᠈పୗࡢཎᅉࢆ⌮ゎࡋࡓ࠸ࠋ ᮏබເᨭㄢ㢟࡛ࡣࠊ⏕యෆࡢ௦ㅰᜏᖖᛶࡢኚࡀࠊ グ᠈ᙧᡂ㛵ࡍࡿศᏊࢿࢵࢺ࣮࣡ࢡ࠼ࡿᙳ㡪 ࡘ࠸࡚ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ㏻ࡌ࡚ホ౯ࡍ ࡿࡇ࡛ࠊ⪁క࠺グ᠈పୗࡀ⏕ࡌࡿ࣓࢝ࢽࢬ࣒ ࡢ⌮ゎ㏕ࡿࡇࢆ┠ⓗࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 2015 ᖺᗘࡢᨭㄢ㢟ࡢࡓࡵࠊࢧࣥࣉࣝ‽ഛࡀ⌧ᅾ㐍 ⾜୰࡛ࡣ࠶ࡿࡀᨭணᐃࡢෆᐜࢆ௨ୗグࡍࠋ ⏕యෆࡢ௦ㅰኚࡼࡾ⬻ෆ࡛Ⓨ⌧ኚࡍࡿ㑇ఏ Ꮚ⩌ࢆྠᐃࡋࡓ࠸ࠋࡲࡓࡑࢀࡽࢆ⪁㱋ಶయ࠾࠸࡚ ࡶẚ㍑ࡍࡿࠋලయⓗࡣࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ 㸦 'URVRSKLOD 0HODQRJDVWHU ࠊ ࢤ ࣀ ࣒ ࢧ ࢬ 㸸 㸧ࡢ L㔝⏕ᆺ࣭ⱝ㱋ಶయ㸦ࣁ࢚ ᪥㱋㸧ࠊ LL㔝⏕ᆺ࣭⪁ಶయ㸦ࣁ࢚ ᪥㱋㸧 ࠊLLL௦ㅰࢆኚ ࡉࡏࡓⱝ㱋ಶయࠊLY௦ㅰࢆኚࡉࡏࡓ⪁㱋ಶయ ࠾ࡅࡿ⬻ 51$ ࢧࣥࣉࣝࢆᅇࡋ࡚ࠊḟୡ௦ࢩ࣮ࢡ࢚ ࣥࢧ࣮㸦51$VHT㸧ࡼࡾ㑇ఏᏊࣉࣟࣇࣜࣥࢢࢆ ẚ㍑ࡍࡿࡇࢆணᐃࡋ࡚࠸ࡿࠋලయⓗࡣࠊ௦ㅰࡢ ኚࡼࡾ⬻ෆ࡛Ⓨ⌧㔞ࡀୖ᪼ࡍࡿࠊࡶࡋࡃࡣపୗ ࡍࡿ㑇ఏᏊ⩌ࢆᢳฟࡋࠊࡑࢀࡽࣜࢫࢺࡘ࠸࡚ⱝ㱋 ಶయ⪁㱋ಶయ࡛ẚ㍑ࡍࡿࠋࡲࡓࠊᚓࡽࢀࡓ㑇ఏᏊ ࣉࣟࣇࣜࣥࢢࡼࡾ *HQH2QWRORJ\ ゎᯒࢆ⾜࠸ࠊ⏕ యෆࡢ௦ㅰᜏᖖᛶࡢኚࡀ⬻ෆࡢ㑇ఏᏊࢿࢵࢺ࣮࣡ ࢡ࠼ࡿᙳ㡪ࢆ᫂ࡽࡍࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⏕యෆࡢ௦ㅰኚࡼࡾᙳ㡪ࡍࡿ⬻ෆ㑇ఏᏊࣉࣟࣇ ࣜࣥࢢࢆẚ㍑ࡍࡿࡇ࡛ࠊ⏕యෆࡢ௦ㅰᜏᖖᛶ ࡀኚࡋࡓ㝿ᙳ㡪ࡍࡿศᏊࢿࢵࢺ࣮࣡ࢡࢆᢳฟࡍ ࡿࡇࡀྍ⬟࡛࠶ࡿ⪃࠼ࡿࠋࡇࢀࡽྠᐃࡋࡓศᏊ ࢿࢵࢺ࣮࣡ࢡࡀᐇ㝿グ᠈ᶵ⬟ᙳ㡪ࡋ࡚࠸ࡿ ࠺ࢆ᫂ࡽࡍࡿࡓࡵࠊศᏊࢿࢵࢺ࣮࣡ࢡෆࡢ 㑇ఏᏊⓎ⌧㔞ࢆᨵኚࡉࡏࡓ⪁㱋ಶయ࠾࠸࡚ࠊႥぬ グ᠈ࢆ⾜ືゎᯒࡼࡾ ᐃࡍࡿࡇࢆணᐃࡋ࡚࠸ࡿࠋ ࡉࡽྠᐃࡉࢀࡓ㑇ఏᏊ⩌ࡘ࠸࡚ࠊLሙᡤ≉␗ ⓗ㑇ఏᏊࢆⓎ⌧ࡉࡏࡿࡇࡀྍ⬟࡞ *DO8$6 㑇ఏ ᏊⓎ⌧⣔࠾ࡼࡧLLᮇ≉␗ⓗ㑇ఏᏊࢆⓎ⌧ࡉ ࡏࡿࡇࡀྍ⬟࡞ *DO8$6 㑇ఏᏊⓎ⌧⣔࡛࠶ࡿ ͆JHQHVZLWFK ࢩࢫࢸ࣒͇㸦5RPDQHWDO31$6㸧 ࢆ⏝࠸࡚ࠊ⚄⤒⣽⬊≉␗ⓗࡘᮇ≉␗ⓗࡑࢀࡒ ࢀࡢ㑇ఏᏊⓎ⌧ࢆᨵኚࡋࡓಶయࢆసᡂࡋࠊࡉࡽࡑ ࢀࡒࢀࡢಶయ࠾ࡅࡿグ᠈⬟ࡢ ᐃࢆ㐍ࡵࡿࡇࢆ ணᐃࡋ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᚋᚓࡽࢀࡿணᐃ࡛࠶ࡿ⤖ᯝࡢゎᯒࡼࡾࠊ⏕యෆ ࡢ௦ㅰᜏᖖᛶࡢኚࡀ࠼ࡿ⬻ෆࡢศᏊࢿࢵࢺ࣮࣡ ࢡࢆ᫂ࡽ࡛ࡁࡿ⪃࠼ࡿࠋࡉࡽࠊグ᠈⥔ᣢ⬟ ࡢ㛵㐃ᛶࡘ࠸᳨࡚ウࡍࡿࡇ࡛ࠊ⏕యෆࡢ௦ㅰ グ᠈ᙧᡂ㛵ࡍࡿศᏊࢿࢵࢺ࣮࣡ࢡࡢ㛵㐃ࡘ࠸ ࡚ࠊᮏᨭࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ㏻ࡌ࡚ホ ౯ࡍࡿࡇࡀ࡛ࡁࡿ⪃࠼ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ‽ഛ୰ - 447 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࣋ࢩࢡࣝࢆࡋࡓᚤ⏕≀㛫ࢿࢵࢺ࣮࣡ࢡࡢゎ᫂ࡑࡢࢹࢨࣥ᪉ἲࡢฟ ⱝᡭ◊✲㸦㸿㸧 ㇏⚟ 㞞࣭⟃ἼᏛ⏕⎔ቃ⣔ 㸺⫼ᬒ࣭┠ⓗ㸼 ⣽⳦ࡣ⣽⬊እ࣓ࣥࣈࣛࣥ࣋ࢩࢡࣝ(MV) ࡤࢀࡿ⭷ᑠ⬊ࢆᨺฟࡍࡿࠋࡇࡢ MV ࡣ㸪⣽⳦㛫ࡸ⣽ ⳦̾ᐟ㛫࡛ࡸࡾࡾࡉࢀࡿࠋᚑࡗ࡚㸪MV ࡢ୰㌟ࡢ ゎᯒࡣࡢࡼ࠺࡞≀㉁ࡀ⣽⬊㛫࡛ఏ㐩ࡉࢀ࠺ࡿࡢ ゎ᫂ࡍࡿୖ࡛㔜せ࡛࠶ࡿࠋࡇࢀࡲ࡛ࡢ◊✲࡛㸪MV ࡣẘ⣲ᅉᏊࡸ⣽⬊㛫ࢩࢢࢼࣝ≀㉁ࡀྵࡲࢀࡿࡇ ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ ࡉࡽࡣ㸪⣽⳦㛫࡛ࡢ㑇ఏᏊࡢỈᖹఏ MV ࡀ 㛵ࡍࡿࡢሗ࿌ࡶ࠶ࡿࠋ⣽⳦࠾ࡅࡿ㑇ఏᏊࡢỈ ᖹఏࡣᩍ⛉᭩ⓗࡣ 1)⣽⬊እ DNA ࢆྲྀࡾ㎸ࡴᙧ ㉁㌿㸪2)ࣇ࣮ࢪࢆࡋࡓᙧ㉁ᑟධ㸪3)᥋ྜఏ㐩, ࡼࡗ࡚⾜ࢃࢀࡿㄝ᫂ࡉࢀ࡚ࡁࡓࠋ MV ࡀ➨ 4 ࡢ 㑇ఏᏊࡢỈᖹఏࡢᢸ࠸ᡭ࡛࠶ࢀࡤ㸪᪤Ꮡࡢྍືᛶ 㑇ఏᏊࡢᴫᛕࡀࡁࡃᢲࡋᗈࡆࡽࢀࡿࡇࡀᮇᚅࡉ ࢀࡿࠋ ᐇ㝿㸪MV DNA ࡀྵࡲࢀ࡚࠸ࡿࡇࡀከᩘࡢ ⣽⳦࡛ሗ࿌ࡉࢀ࡚࠸ࡿࡀ㸪ࡑࡢࢩ࣮ࢡ࢚ࣥࢫࡘ࠸ ࡚ゎᯒࡋࡓࡣࢇ࡞࠸ࠋࡑࡇ࡛ᮏㄢ㢟࡛ࡣ㸪 MV ྵࡲࢀࡿ DNA ࢩ࣮ࢡ࢚ࣥࢫࢆ᫂ࡽࡍࡿ ࡇࡼࡗ࡚㸪ࡢࡼ࠺࡞㑇ఏᏊࡀ MV ྵࡲࢀ࡚ ࠾ࡾ㸪Ỉᖹఏࡋ࠺ࡿࡢ᫂ࡽࡍࡿࡇࢆ┠ⓗ ࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᮏ ᨭ ࡛ ࡣ 㸪 ࢢ ࣛ ࣒ 㝜 ᛶ ⣽ ⳦ ࡢ Burkholderia cenocepacia H111 ཬࡧ Paracoccus denitrificans Pd1222 ࢢ࣒ࣛ㝧ᛶ⣽⳦ࡢ Bacillus subtilis 168 ཬࡧ Clostridium perfringens 13 ࡢ MV ྵࡲࢀࡿ DNA( ௨ ୗ mvDNA) ࡘ ࠸ ࡚ ゎ ᯒ ࡋ ࡓ ࠋ ࡲ ࡓ 㸪 mvDNA 㓄ิࡀ⎔ቃࡸ MV ᙧᡂᶵᵓ౫Ꮡⓗゎᯒࡍ ࡿࡓࡵ㸪B. cenocepacia H111 ཬࡧ B. subtilis 168 ࡘ࠸࡚ࡣ㸪✀㢮ࡢ᮲௳ୗ࡛ᅇࡋࡓ MV ࡢ mvDNA ࢆࢩ࣮ࢣࣥࢩࣥࢢࡋࡓࠋୖグࡢィ 6 ࢧࣥࣉ ࣝࢆᮏᨭ࡛ࢩ࣮ࢣࣥࢩࣥࢢࡋ㸪ࡉࡽࡣᮏᨭ࡛㸪 ᚓࡽࢀࡓࢩ࣮ࢣࣥࢫࢆࢤࣀ࣒ሗᑐࡋ࡚࣐ࢵࣆࣥ ࢢࡋࡓࠋ࣐ࢵࣆࣥࢢࡢ⤖ᯝ㸪ࡢࡼ࠺࡞㑇ఏᏊࡀ MV ྵࡲࢀ㸪 ࡑࡢ㓄ิ೫ࡾࡀ࠶ࡿ࠺, FPKM ್ ࢆィ⟬ࡍࡿࡇ࡛ゎᯒࢆ⾜ࡗࡓࠋᮏᨭࡼࡗ࡚ࡍ ࡛ 6 ࢧࣥࣉࣝࡍ࡚ࡢࢩ࣮ࢣࣥࢩࣥࢢࡀࡋ࡚ ࠸ ࡿ ࠋࡑ ࡢ࠺ ࡕ㸪 P. denitrificans Pd1222 B. subtilis 168 ࡘ࠸࡚ࡣ㸪࣐ࢵࣆࣥࢢࡶࡋ㸪ṧࡾ ࡢᰴࡘ࠸࡚ࡶᘬࡁ⥆ࡁ࣐ࢵࣆࣥࢢࢆ⾜ࡗ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⏦ㄳ⪅ࡣ Pseudomonas aeruginosa ࢆ⏝࠸ࡓඛ⾜ ◊✲࡛㸪 MV ᙧᡂ㛵ࡍࡿ㑇ఏᏊࢆぢฟࡋ࡚࠸ࡿ (Nature communications ࣜ ࣂ ࢫ ୰ ) ࠋ P. denitrificans Pd1222 ࡘ࠸࡚㸪ࢤࣀ࣒㡿ᇦࡀ MV ྵࡲࢀ࡚࠸ࡿࡶࡢࡢ㸪⯆῝࠸ࡇ㸪ࡑࡢ࠺ࡕ ࡢ≉ᐃࡢ 㡿ᇦࡀ㠀ᖖከࡃ✚ࡉࢀ࡚࠸ࡿࡇࡀ ☜ㄆࡉࢀࡓࠋᮏ㡿ᇦࡣ P. aeruginosa ࡢ MV ᙧᡂ 㛵ࢃࡿ㑇ఏᏊࡢ࣍ࣔࣟࢢࡶྵࢇ࡛࠾ࡾ㸪ྠᵝࡢᶵᵓ ࡛ MV ࡀᨺฟࡉࢀ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿࠋ୍᪉࡛㸪B. subtilis 168 ࡘ࠸࡚ࡣࢤࣀ࣒㡿ᇦࡀࡰ➼㔞ྵ ࡲࢀ࡚࠸ࡓࠋࡇࡢࡇࡣ㸪DNA ࡣࣛࣥࢲ࣒ MV ྵ ࡲࢀࡿࡇࢆ♧၀ࡍࡿࠋB. subtilis 168 ࡢ MV ᙧᡂ ᅉᏊࢆ⌧ᅾྠᐃࡋ࡚࠾ࡾ㸪ᙜヱ MV ᙧᡂᶵᵓࢆ㋃ࡲ ࠼ࡿ㸪ጇᙜ࡞⤖ᯝ࡞ࡗࡓࠋࡉࡽ㸪⎔ቃ᮲௳ࡢ 㐪࠸࡛ MV ྵࡲࢀࡿ DNA 㓄ิࡁ࡞ᕪ␗ࡀほ ᐹࡉࢀࡿ࠺ゎᯒ୰࡛࠶ࡿࠋ௨ୖ㸪ᮏᨭࡼ ࡗ࡚ MV ྵࡲࢀࡿ DNA 㓄ิࡀ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 MV ྵࡲࢀࡿ㑇ఏᏊࡣ⣽⳦㛫࡛Ỉᖹఏࡍࡿࡇ ࡀ▱ࡽࢀ࡚࠸ࡓࡀ㸪ᐇ㝿 MV ࡢࡼ࠺࡞ DNA ࡀྵࡲࢀ࡚࠸ࡿࡢࡘ࠸࡚ࡑࡢヲ⣽ࡣ᫂ࡽ࡞ ࡗ࡚࠸࡞ࡗࡓࠋ ᮏᨭࡼࡗ࡚㸪MV ྵࡲࢀࡿ DNA 㓄ิࡀ᫂ࡽ ࡞ࡾ㸪Ỉᖹఏࡋ࠺ࡿ㑇ఏᏊࢆ⌮ゎࡍࡿୖ࡛㔜 せ࡞▱ぢࡀᚓࡽࢀࡓࠋࢤࣀ࣒ሗࡀ✚ࡉࢀ࡚࠸ࡃ ࡘࢀ㸪⣽⳦ࡢࢤࣀ࣒⦅ᡂỈᖹఏࡀ࠸㛵ࢃ ࡗ࡚࠸ࡿࡀ᫂ࡽ࡞ࡗ࡚ࡁ࡚࠸ࡿࡀ㸪ࡢࡼ࠺ 㑇ఏᏊࡀఏࡉࢀࡓࡢࡘ࠸࡚ࡣ㸪ࡣ⌮ ゎࡉࢀ࡚࠸࡞࠸ࠋᮏᨭᡂᯝࡣ MV ࡀ㑇ఏᏊࡢỈᖹ ఏࡢᢸ࠸ᡭࡋ࡚㔜せ࡛࠶ࡿࡇࢆ♧၀ࡍࡿࠋ ᭱⤊ⓗ㑇ఏᏊࡀᐟྲྀࡾ㎸ࡲࢀࡿࡓࡵࡣ㸪 DNA ࡢไ㝈⣔ࡸ┦ྠ⤌࠼࡞㸪ࡢせᅉࡀᅾࡍ ࡿࡀ㸪ᮏゎᯒࡼࡿ㸪ࢤࣀ࣒ࡢከᩘࡣ⣽⬊㛫࡛ ఏࡍࡿྍ⬟ᛶࡀ࠶ࡿࠋᚋ㸪㑇ఏᏊ࣐࣮࣮࢝࡞ ࢆ⏝࠸࡚㸪ᐇ㝿ࡢࢤࣀ࣒㡿ᇦࡀỈᖹఏࡍࡿࡢ 㸪ࡲࡓࡑࡢ㝿ࡢ㝈ᐃᅉᏊࡣఱ࡛࠶ࡿࡢ➼ゎᯒࡍ ࡿࡓࡵࡢᇶ♏ⓗ࡞▱ぢࡀᚓࡽࢀࡓࠋ MV ࡢ୰㌟ࢆㄪࡿࡇࡣ㸪MV ࡀࡢࡼ࠺ᙧ ᡂࡉࢀࡓࡘ࠸࡚ࡶ㔜せ࡞♧၀ࢆ࠼ࡿࠋᮏᡂᯝ ࡛ mvDNA 㓄ิࡀ᫂ࡽ࡞ࡗࡓࡇ࡛, MV ᙧᡂ 㛵ࢃࡿᅉᏊࢆ⤠ࡾ㎸ࡴࡇࡀ࡛ࡁࡓࠋᚋ㑇ఏᏊ Ḟᦆᰴࡢస〇ࢆ㏻ࡌ࡚㸪MV ᙧᡂ㛵ࢃࡿ㑇ఏᏊࢆ ྠᐃࡍࡿணᐃ࡛࠶ࡿࠋMV ᙧᡂᶵᵓࡣࡲࡔゎ ᫂ࡉࢀ࡚࠾ࡽࡎ㸪MV ࡢ⏕≀Ꮫⓗ࡞ᙺࢆ⪃࠼ࡿ㸪 ࡑࡢᙧᡂᶵᵓࡢゎ᫂ࡣࡁ࡞ㄢ㢟࡛࠶ࡿࠋࡉࡽ㸪 MV ࡣ࣡ࢡࢳࣥ࡞ࡶ⏝ࡉࢀ࡚࠾ࡾ㸪㑇ఏᏊ⤌ ࠼ᢏ⾡ࢆ㐺⏝ࡋࡸࡍ࠸ࡇࡶ࠶ࡗ࡚㸪MV ࡀ┒ࢇ ᛂ⏝ࡉࢀࡘࡘ࠶ࡿࠋMV ࢆ㔞ᥦ౪࡛ࡁࡿࡀ ᚋࡢㄢ㢟࡞ࡿࡀ㸪MV ᙧᡂᶵᵓࡢゎ᫂ࡣ MV ࡢ 㔞⏕⏘⧅ࡀࡿྍ⬟ᛶࡀ࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 448 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࢭࣥࢺ࣓ࣟ⦅ࡢศᏊ࣓࢝ࢽࢬ࣒ ᪂Ꮫ⾡㡿ᇦ◊✲ ୰ᕝ ᣅ㑻࣭㜰ᏛᏛ㝔⌮Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ᰁⰍయࡢ␗ᖖࡼࡾ⒴࡞ࡢ㑇ఏࡀᘬࡁ㉳ࡇࡉ ࢀࡿࠋᰁⰍయࡢࢭࣥࢺ࣓ࣟ㡿ᇦࡣṇ☜࡞ᰁⰍయศ 㓄㔜せ࡛࠶ࡿࠋ⯆῝࠸ࡇࠊࣄࢺࢆྵࡴከࡃ ࡢ┿᰾⏕≀ࡢࢭࣥࢺ࣓ࣟࡣࣜࣆ࣮ࢺ㓄ิࡀᏑᅾ ࡍࡿࡓࡵࠊྠ⭎ᰁⰍయᙧᡂࡸࣟࣂ࣮ࢺࢯࣥ㌿ᗙ࡞ ࡢᰁⰍయ⦅ࡀ㉳ࡇࡿࠋࡇࢀࡲ࡛ࡢ◊✲ࡽࠊDNA ┦ྠ⤌࠼ࡸ⣽⬊࿘ᮇࢳ࢙ࢵࢡ࣏ࣥࢺ࡞ࡀᰁⰍ య⦅ࡢᢚไ㔜せ࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ࡋࡋࠊᰁⰍయ⦅ࡑࡢࡶࡢࡢศᏊ࣓࢝ࢽࢬ࣒ࡣ࠸ ࡲࡔゎ᫂ࡉࢀ࡚࠸࡞࠸ࠋ ࡑࡇ࡛ࠊᡃࠎࡣศ㓝ẕ Schizosaccharomyces pombe ࢆ⏝࠸࡚ࢭࣥࢺ࣓࡛ࣟࡢᰁⰍయ⦅ᚲせ ࡞ ᅉ Ꮚ Factors of Centromere rearrangement (FOC)ࢆྠᐃࡋࠊFOC ࡢᙺᶵ⬟ࢆ᫂ࡽࡍࡿࠋ ᭦ࠊ⢭〇 FOC ⺮ⓑ㉁ࢆ⏝࠸࡚ヨ㦂⟶ෆ࡛ࢭࣥࢺࣟ ࣓⦅ࢆᵓᡂࡍࡿࡇ࡛ࠊࢭࣥࢺ࣓ࣟ⦅ࡢ ศᏊ࣓࢝ࢽࢬ࣒㏕ࡿࠋᮏ◊✲ࡼࡾᚓࡽࢀࡿᡂᯝ ࡣࠊࢭࣥࢺ࣓ࣟ㝈ࡽࡎࢺࣛࣥࢫ࣏ࢰࣥ࡞⏤ ᮶ࡍࡿࣜࣆ࣮ࢺ㛫࡛㉳ࡁࡿᰁⰍయ⦅ࡢᬑ㐢ⓗ࡞࣓ ࢝ࢽࢬ࣒ࡢ⌮ゎᙺ❧ࡘ⪃࠼ࡽࢀࡿࠋࡲࡓࠊࡀࢇ ࡢⓎࢆண㜵ࡍࡿ⸆ࡢ㛤Ⓨࡶ㈉⊩ࡍࡿ⪃࠼ࡽ ࢀࡿࠋ 㸺ᨭࡢෆᐜ㸼 ศ㓝ẕ S. pombe ࡢ᪂つኚ␗ᰴࢥࣥࢺ࣮ࣟࣝ ᰴࡢࢤࣀ࣒ DNA ࢆᯘဴஓඛ⏕㸦ᕞᏛ㸧ࡢࡈ༠ຊ ࡼࡾࣜࢩ࣮ࢣࣥࢫࡋ࡚㡬࠸ࡓࠋࡑࡋ࡚ࠊࡑࢀࡽࡢ 㛫ࡢሷᇶ㓄ิẚ㍑ࡍࡿࡇ࡛ࠊኚ␗㒊ࢆỴᐃࡋ ࡓࠋ௨ୗࡑࡢヲ⣽ࢆㄝ᫂ࡍࡿࠋ 㸦㸯㸧ࢤࣀ࣒ DNA ࡣ Positive pool Negative pool ࡢ㸰ࡘࡢࢧࣥࣉࣝࢆ㸯ࢭࢵࢺࠋࡇࢀࢆ㸱ࢭࢵࢺ㔝 ⏕ᰴࡢࢤࣀ࣒ DNA ࢆྜࢃࡏ࡚ィ㸵ࢧࣥࣉࣝࡢ㓝ẕ ᰴࡢࢤࣀ࣒ DNA ࢆ MiSeq㸦Illumina ♫㸧ࢆ⏝࠸࡚ ࢚࣌ࣥࢻἲࡼࡾࣜࢩ࣮ࢡ࢚ࣥࢫࡋ࡚㡬࠸ࡓࠋ⣽ ⬊ࢆள◪㓟࡛ฎ⌮ࡋࡓࡓࡵᩘከࡃࡢኚ␗ࡀࢤࣀ࣒ DNA ᑟධࡉࢀ࡚࠸ࡓࠋࡑࡇ࡛ࠊPooled Linkage Analysis (PLA)ࡤࢀࡿ᪉ἲࢆ᥇⏝ࡋࡓࠋPLA ἲ ࡛ࡣ⊂❧ࡋࡓኚ␗ᰴࡢࢤࣀ࣒ DNA ࢆࣉ࣮ࣝࡋࡓᚋ ࣜࢩ࣮ࢣࣥࢫࢆ⾜࠺ࡓࡵࠊẚ㍑ⓗከࡃࡢ㔜」㸦100x coverage㸧࡛ሷᇶ㓄ิࡢゎㄞࢆ⾜ࡗ࡚㡬࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᅇࠊࣜࢩ࣮ࢡ࢚ࣥࢫࣂ࢜ࣥࣇ࢛࣐ࢸࢵ ࢡࢫࢆࡈᨭ㡬࠸ࡓࡇࡼࡾࠊ┠ⓗࡋ࡚࠸ࡓᰁ Ⰽయ⦅ᚲせ࡞᪂つ㑇ఏᏊ FOC1 ࡢྠᐃᡂຌࡋ ࡓࠋFOC1 ࡢᙺࢆヲࡋࡃゎᯒࡍࡿⅭ㑇ఏᏊ◚ቯ ᰴࢆసᡂࡋࡓ⤖ᯝࠊ⯆῝࠸ࡇࠊFOC1 ࡣ DNA ┦ྠ⤌࠼ᅉᏊ Rad51 ࡢ◚ቯᰴ࡛㉳ࡁࡿᰁⰍయ⦅ ≉㔜せ࡛࠶ࡿࡇࡀศࡗࡓࠋࡲࡓࠊfoc1 ◚ቯ ᰴࡣ DNA ࣝ࢟ࣝ㸦࣓ࢳ࣓ࣝࢱࣥࢫࣝ࣍ࣥ㓟 MMS㸧࡞ᑐࡋ࡚㧗࠸ឤཷᛶࢆ♧ࡍࡇࠊࡑࡋ࡚ࠊ ࡑࡢ DNA ࢲ࣓࣮ࢪឤཷᛶࡣ rad51 ࢆ◚ቯࡍࡿࡇ ࡛ࡼࡾ㧗ࡃ࡞ࡿࡇࡀศࡗࡓࠋࡇࢀࡽࡢ⤖ᯝࡽࠊ DNA ⤌࠼ಟ࠸࡚ Rad51 ࡣ⊂❧ࠊFoc1 ࡣാࡁࠊᰁⰍయ⦅ࢆᘬࡁ㉳ࡇࡍᅉᏊ࡛࠶ࡿ⪃࠼ ࡽࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᅇࠊศ㓝ẕࡢ᪂つኚ␗ᰴࡢࢫࢡ࣮ࣜࢽࣥࢢ ḟୡ௦ࢩ࣮ࢣࣥࢫࢆᑟධࡋࡓࠋࡇࡢࡈᨭࡼࡾࠊ ᚑ᮶ࡢᡭἲ࡛ࡣᅔ㞴࡞ࢣ࣮ࢫࠊ࠶ࡿ࠸ࡣࠊつᶍࢫ ࢡ࣮ࣜࢽࣥࢢࢆ⾜࠺ࢣ࣮ࢫ࡞࡛ࡣࠊḟୡ௦ࢩ࣮ࢣ ࣥࢫࢆά⏝ࡍࡿࡇࡀ㠀ᖖ᭷ຠ࡛࠶ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋᚋࡢ◊✲ࡶḟୡ௦ࢩ࣮ࢣࣥࢫࢆά ⏝ࡍࡿ㐨ࡍࡌࡀ㛤ࢀࡓࠋ ࣄࢺࡢሙྜࠊBRCA2 ࡞ࡢ┦ྠ⤌࠼ᅉᏊኚ␗ ࡀ࠶ࡿ␗ᖖᰁⰍయࡀ㧗㢖ᗘ࡛ᙧᡂࡉࢀࡿࡇ࡛ங ࡀࢇ࡞ࡢࡀࢇࡀㄏⓎࡉࢀࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋ ᅇࡢࢫࢡ࣮ࣜࢽࣥࢢࡼࡾྠᐃࡋࡓ FOC㸯ࡣ㓝ẕ ࡽࣄࢺ⮳ࡿࡲ࡛ᗈࡃ┿᰾⏕≀ಖᏑࡉࢀ࡚࠸ࡿࠋ ࡋࡓࡀࡗ࡚ࠊᚋࠊᇵ㣴⣽⬊ࢆ⏝࠸࡚ࠊFOC1 ࡢᙺ ࡸᶵ⬟ࢆゎᯒࡍࡿࡇ࡛ࣄࢺࡅࡿᰁⰍయ⦅ ࡢ࣓࢝ࢽࢬ࣒ࡶ᫂ࡽ࡞ࡿᮇᚅࡋ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᢞ✏ㄽᩥ‽ഛ୰ 㸦㸰㸧ࣜࢩ࣮ࢣࣥࢫࡼࡾᚓࡽࢀࡓሷᇶ㓄ิሗࢆ බ㛤ࡉࢀ࡚࠸ࡿศ㓝ẕࡢࢤࣀ࣒㓄ิ࣐ࢵࣆࣥࢢ ࡍࡿࠋࡲࡓࠊPositive pool Negative pool ࡛ẚ㍑ ゎᯒࡋ࡚ࠊኚ␗㒊ࢆ≉ᐃࡍࡿࣂ࢜ࣇ࢛࣐ࢸ ࢵࢡࢫゎᯒࡶࡈᨭ㡬࠸ࡓࠋ - 449 - බເᨭㄢ㢟㸸2012 ᖺᗘ ேⅭ㉳※≀㉁ศゎ⬟ࢆࣔࢹࣝࡋࡓ⎔ቃ୰࡛ࡢ⣽⳦ࡢᶵ⬟㐍ࢆྖࡿศᏊᇶ┙ࡢゎ᫂ ᇶ┙◊✲㸦㹀㸧 Ọ⏣ ⿱࣭ᮾᏛᏛ㝔⏕⛉Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ௦⾲ⓗ࡞⎔ቃởᰁ≀㉁࡛࠶ࡿ᭷ᶵሷ⣲⣔ẅ J-hexachlorocyclohexane (J-HCH)ࡣᏛྜᡂࡼࡾ ⏕⏘ࡉࢀࡓᆺⓗ࡞ேⅭ㉳※≀㉁࡛࠶ࡾࠊJ-HCH ศゎ⣽⳦ࡀ⎔ቃ୰࡛ዴఱࡋ࡚ฟ⌧ࡋࡓࢆࠕࢤ ࣀ࣒ࠖࠕศゎ㓝⣲㑇ఏᏊࠖ╔┠ࡋ࡚ゎ᫂ࡍࡿ ◊✲ࢆࠕேⅭ㉳※≀㉁ศゎ⬟ࢆࣔࢹࣝࡋࡓ⎔ቃ ୰࡛ࡢ⣽⳦ࡢᶵ⬟㐍ࢆྖࡿศᏊᇶ┙ࡢゎ᫂ࠖ 㸦ᇶ ┙◊✲(B) ᖹᡂ 22~24 ᖺᗘ㸧࡛ᐇࡋࡓࠋࡑࡢ㐣 ⛬࡛ࠊᐇ㝿ࡢ⎔ቃ୰࡛ࡣࠊ༢⊂ࡢศゎ⣽⳦࡛ࡣ࡞ ࡃࠊ」ᩘࡢ⣽⳦✀ࡽ࡞ࡿࢥ࣑ࣗࢽࢸࡼࡗ࡚ J-HCHࡀ௦ㅰࡉࢀ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋࡍ ࡞ࢃࡕࠊ᪥ᮏࡢ HCH ␗ᛶయởᰁᅵተヨᩱࡽࠊ J-HCH ࢆ၏୍ࡢⅣ⣲※ࡋࡓ↓ᶵᾮయᇵᆅ࡛㞟✚ ᇵ㣴ࢆ⾜ࡗࡓࡇࢁࠊ」ᩘࡢ⣽⳦✀ࡽ࡞ࡿࢥ࣑ ࣗࢽࢸࡀ㞟✚ࡉࢀࠊᾮయᇵᆅࢆ⏝࠸ࡓẁ㝵ᕼ㔘 ἲ࡛ࡶ༢⊂ࡢศゎ⣽⳦ࢆศ㞳ࡍࡿࡇࡀ࡛ࡁ࡞ ࡗࡓࠋࡉࡽࠊᮏ⣽⳦ࢥ࣑ࣗࢽࢸࡽᖹᯈᕼ㔘 ἲࢆ⧞ࡾ㏉ࡍࡇ࡛ࠊJ-HCH ࢆ၏୍ࡢⅣ⣲※ࡋ ࡚⏕⫱ࡍࡿ Sphingobium sp. TKS ᰴJ-HCH ࡢศゎ άᛶࢆ᭷ࡉ࡞࠸ Pseudomonas sp. TKP ᰴࢆ༢㞳ࡋ ࡓࠋ୍᪉ࠊࣥࢻࡢ HCH ␗ᛶయởᰁᅵተヨᩱࡽࠊ t-HCH (D-, E-, G-, J-HCH ࡢΰྜ≀)ࢆྵࡴపᰤ㣴ᾮ యᇵᆅࡼࡿ㞟✚࡛ᚓࡽࢀࡓ⣽⳦ࢥ࣑ࣗࢽࢸ ࡽࡶࠊྠᵝJ-HCH ࢆ၏୍ࡢⅣ⣲※ࡋ࡚⏕⫱ࡍ ࡿ Sphingomonas sp. MM-1 ᰴJ-HCH ࡢศゎάᛶ ࢆ᭷ࡉ࡞࠸ Pseudomonas aeruginosa MTB-1 ᰴࢆ༢ 㞳ࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ௨ୖࡢ⫼ᬒࡢඖࠊJ-HCH ศゎ⣽⳦ࢥ࣑ࣗࢽࢸ ࡢᵓ㐀ᶵ⬟ࡢゎ᫂ࢆ┠ⓗࡋ࡚ࠊ௨ୗࡢ㸱㡯┠ ࡢࢤࣀ࣒ᨭࢆཷࡅࡓࠋࢤࣀ࣒ᨭ࠾ࡅࡿ 454 ࠾ࡼࡧ illumina ࡼࡿゎᯒࡽᚓࡽࢀࡓ࣮ࣜࢻࢹ ࣮ࢱࡢᴫせࢆేࡏ࡚グࡍࠋձ ᪥ᮏࡢ HCH ␗ᛶయ ởᰁᅵተヨᩱ⏤᮶ࡢJ-HCH ศゎ⣽⳦ࢥ࣑ࣗࢽࢸ ࡢ࣓ࢱࢤࣀ࣒ゎᯒ 㸦total read ࡋ࡚ࠊ454 ࡛ 1 Gb ⛬ ᗘ ࠊ illumina ࡛ 5.5 Gb ⛬ ᗘ 㸧 ղ ༢ 㞳 ࡋ ࡓ Pseudomonas sp. TKP ᰴࡢࢤࣀ࣒ゎᯒ㸦total read ࡋ࡚ࠊ454 ࡛ 240 Mb ⛬ᗘࠊillumina ࡛ 50 Mb ⛬ᗘ㸧 ճ ༢㞳ࡋࡓ Pseudomonas aeruginosa MTB-1 ᰴࡢ ࢤࣀ࣒ゎᯒ㸦total read ࡋ࡚ࠊ454 ࡛ 200 Mb ⛬ᗘࠊ illumina ࡛ 500 Mb ⛬ᗘ㸧 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᨭࢹ࣮ࢱࢆඖ TKP ᰴ MTB-1 ᰴࡢࢤࣀ ࣒㓄ิࢆỴᐃࡋࠊㄽᩥⓎ⾲ࡋࡓࠋMTB-1 ᰴࡣ Pseudomonas aeruginosa ᰴ᭱ࡶ㏆⦕࡛࠶ࡾࠊࢹ࣮ ࢱ࣮࣋ࢫୖࡢ P. aeruginosa ࢤࣀ࣒Ỵᐃᰴ యⓗ㧗࠸㢮ఝᛶࢆ♧ࡋࡓࠋMTB-1 ᰴ≉␗ⓗ࡞㡿 ᇦࡣ phage/island ᥎ᐃࡉࢀࡓࠋ୍᪉ࠊTKP ᰴࡣ P. fluorescens SWB ᰴࡸ P. poae RE*1-1-14 ᰴẚ㍑ ⓗ㧗࠸㢮ఝᛶࢆ♧ࡋࡓࡀࠊ✀࡛ࣞ࣋ࣝࡣ≉ᐃ࡛ࡁ ࡎࠊሗࡢᑡ࡞࠸ Pseudomonas ᒓᰴࡢࢤࣀ࣒㓄 ิࢆỴᐃ࡛ࡁࡓࠋࡇࢀࡽ 2 ᰴࡣ⌮◊ JCM ࡛ධᡭ࡛ ࡁࠊࣂ࢜ࣜࢯ࣮ࢫࡋ୍࡚⯡⏝ྍ⬟࡛࠶ࡿࠋ ࡲࡓࠊMTB-1 ᰴࠊTKP ᰴඹJ-HCH ௦ㅰ㛵ࡍ ࡿ lin 㑇ఏᏊ⩌ࠊ࠾ࡼࡧJ-HCH ௦ㅰ୰㛫యࡢ௦ㅰ 㛵ࡍࡿ᥎ᐃࡉࢀࡿ㑇ఏᏊ⩌ࡣぢฟࡉࢀ࡞ࡗ ࡓࠋ୍᪉ࠊ࣓ࢱࢤࣀ࣒ゎᯒࡼࡾࠊJ-HCH ศゎ⣽ ⳦ࢥ࣑ࣗࢽࢸࡣࠊTKS ᰴ TKP ᰴ௨እࡶᒓ ࡛ࣞ࣋ࣝ␗࡞ࡿ」ᩘᰴࡀᏑᅾࡋ࡚࠸ࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋCupriavidus (E-proteobacteria) ࡢྜ ࡀ᭱ࡶ㧗ࡃࠊࡑࢀ Sphingobium (D-proteobacteria)ࠊ Pseudomonas (J-proteobacteria) ࡀ⥆ࡃ࠸࠺⤖ᯝ ࡞ࡗࡓࠋ௨ୖࡢ⤖ᯝࡽࠊࢥ࣑ࣗࢽࢸࡢ㑇ఏ ⓗᵓ㐀㛵ࡍࡿሗࢆᚓࡿඹࠊࡑࡢᙧᡂព⩏ ࡋ࡚ࠊJ-HCH ศゎ௦ㅰ࠾ࡅࡿࢥ࣑ࣗࢽࢸᵓ ᡂ⣽⳦ᰴࡼࡿ✚ᴟⓗ࡞௦ㅰࡢศᴗࡢྍ⬟ᛶࡣప ࠸ࡢ⤖ㄽ⮳ࡗࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࣓ࢱࢤࣀ࣒࣮ࣜࢻࢹ࣮ࢱࡽ࡞ࢥ࣑ࣗࢽࢸ ᵓᡂ⳦ᰴ⪃࠼ࡽࢀࡿ Cupriavidus sp. TKC ᰴࠊ Achromobacter sp. TKA ᰴࠊ Hyphomicrobium sp. TKH ᰴࡢ༢㞳ᡂຌࡋࡓࠋJ-HCH ศゎ⣽⳦ࢥ࣑ࣗࢽࢸ ࡣࠊJ-HCH ࢆ၏୍ࡢⅣ⣲※ࡋ࡚ྵࡴ↓ᶵᐮኳ ᇵᆅୖ࡛ࠊTKS ᰴ༢⊂ࡢሙྜࡼࡾ㛗ᮇ㛫Ώࡗ࡚ ᣢ⥆ⓗJ-HCH ࢆศゎྍ⬟࡞ᕧࢥࣟࢽ࣮ࢆᙧᡂ ࡍࡿሙྜࡀ࠶ࡾࠊࡇࢀࡽ༢㞳⣽⳦ᰴࡀࡇࡢࡼ࠺࡞ ࢥࣟࢽ࣮ᙧᡂ㛵ࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋ ᚋࠊࡇࢀࡽ༢㞳⣽⳦ᰴᮏࢤࣀ࣒ᨭ࡛ᚓࡽࢀ ࡓሗࢆ⏝ࡋ࡚ࠊJ-HCH ศゎ⣽⳦ࢥ࣑ࣗࢽࢸ ࡢᙧᡂཎ⌮ࡢゎ᫂ࠊ࠾ࡼࡧ」ᩘ⣽⳦ᰴࡽ࡞ࡿࢥ ࣟࢽ࣮ࡢ✵ⓗᙧᡂ㐣⛬ࡑࡢᶵᵓࡢゎ᫂◊ ✲ࢆᒎ㛤ࡋ࡚࠸ࡃணᐃ࡛࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ㄽᩥⓎ⾲ Ohtsubo Y et al. 2014. Complete genome sequence of Pseudomonas aeruginosa MTB-1, isolated from a microbial community enriched by the technical formulation of hexachlorocyclohexane. Genome Announcements 2:e01130-13. Ohtsubo Y et al.. 2014. Complete genome sequence of Pseudomonas sp. strain TKP, isolated from a J-hexachlorocyclohexane-degrading mixed culture. Genome Announcements 2:e01241-13. ࢹ࣮ࢱ࣮࣋ࢫⓏ㘓 Pseudomonas aeruginosa MTB-1 ᰴ (JCM 19686) ࢤࣀ࣒㓄ิ Acc No CP006853 බ㛤 Pseudomonas sp. TKP ᰴ (JCM 19688) ࢤࣀ࣒㓄ิ Acc No CP006852 බ㛤 - 450 - බເᨭㄢ㢟㸸2011 ᖺᗘ ࣈ࣮ࣝࣜ₽⒆ᑐ⟇᭷⏝࡞᪩ᮇデ᩿ἲཬࡧ⒪ἲ㛤Ⓨ㛵ࡍࡿ◊✲ ᇶ┙◊✲㸦㹁㸧 ୰Ọ ᯞ࣭ᅜ❧ឤᰁ◊✲ᡤࣁࣥࢭࣥ◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ࣈ࣮ࣝࣜ₽⒆ࡣ Mycobacterium ulcerans ឤᰁࡼ ࡗ࡚⏕ࡌࡿቯṚᛶࡢ㞴ᛶ₽⒆ࢆ≉ᚩࡍࡿ⓶ ᝈ࡛ࠊࡋ࡚⇕ᖏ࣭ள⇕ᖏᆅᇦ࡛ぢࡽࢀࡿࠋWHO ࡢᐃࡵࡓ 17 ᝈࠊNeglected Tropical Diseases ࡢࡦ ࡘࡋ࡚ࠊࡑࡢᑐ⟇ࡀ㐍ࡵࡽࢀ࡚࠾ࡾࠊୡ⏺࡛ẖ ᖺᩘ༓ேつᶍࡢ᪂つᝈ⪅ࡀⓏ㘓ࡉࢀ࡚࠸ࡿࠋᮏ㑥࡛ ࡣ 1980 ᖺ M. ulcerans ㏆⦕ࡢ ͇M. ulcerans subsp. shinshuense͇ (M. shinshuense) ឤᰁࡼࡿ⓶ ₽⒆ࡢ୍ࡀึࡵ࡚ሗ࿌ࡉࢀࡓࠋ M. ulcerans ࡢཎᛶࡢయࡣࡢᢠ㓟⳦✀ࡣ ࡳࡽࢀ࡞࠸ࣘࢽ࣮ࢡ࡞࣐ࢡࣟࣛࢻ mycolactone ࡼࡿࡶࡢ࡛ࠊ⣽⬊Ṛࡸච⣽⬊ࡢᢚไࠊ③ࡳࡢᢚไ ࡞ࢆᘬࡁ㉳ࡇࡍࡶࡢ⪃࠼ࡽࢀ࡚࠸ࡿࠋࡇࡢ mycolactone ࢆ⏘⏕ࡍࡿ⏕ྜᡂ㓝⣲⩌ࡣࡍ࡚⣙ 170kbp ࡢᕧ࡞ plasmid code ࡉࢀࡓᙧ୍࡛⣽⬊ ࠶ࡓࡾ 1-2 ࢥࣆ࣮ࡀᏑᅾࡍࡿࠋM. ulcerans ࡢࢤࣀ࣒ ◊✲ࡣࠊ᭱ึ࣮࢞ࢼࡢࣈ࣮ࣝࣜ₽⒆⨯ᝈ⪅ࡼࡾศ 㞳(1999 ᖺ)ࡉࢀࡓ Agy99 ᰴࡘ࠸࡚ࠊࢤࣀ࣒ࡢヲ ⣽࡞ゎᯒ⤖ᯝࡀሗ࿌ࡉࢀࠊࡑࡢᚋከࡃࡢᰴࡘ࠸࡚ ࢡࣟࣔࢰ࣮࣒㑇ఏᏊࡀẚ㍑ゎᯒࡉࢀ࡚ࡁࡓࠋࡋࡋࠊ ࡇࢀࡲ࡛ࡢሗ࿌࡛ࡣ M. shinshuense ࡢࡘ࠸࡚ࡢ ヲ⣽ࡣ᫂░࡞ࡲࡲ࡛࠶ࡿࠋᮏ㑥ࡢࣈ࣮ࣝࣜ₽⒆ࡣ 2015 ᖺ 12 ᭶ᮎࡲ࡛ィ 57 ࡀ☜ㄆࡉࢀᩘࡣ ᖺࠎቑຍࡋࠊM. shinshuense ࡢࢤࣀ࣒ゎᯒࡢᚲせᛶ ࡣቑࡋ࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᪥ ᮏ ࡛ ᭱ ึ ศ 㞳 ࡉ ࢀ ࡓ ͆ Mycobacterium ulcerans subsp. shinshuense͇ATCC 33728 ᰴ (ẘᛶ⬡㉁ mycolactone ࢆ⏘⏕ࡋ㯤Ⰽࢥࣟࢽ࣮ࠊ ShT-P ᰴ㸧࠾ࡼࡧࠊATCC 33728 ᰴࡼࡾᚓࡽࢀࡓࢥ ࣟࢽ࣮ࡢⰍㄪ࠾ࡼࡧཎᛶࡢ␗࡞ࡿᰴ㸦࣮࣎ࣜ Ⰽࡢࢡ࣮ࣟࣥ ShT-N㸧ࡢィ 2 ᰴࡘ࠸࡚ࡢࢤࣀ࣒ ゎᯒ࠾ࡼࡧ᪥ᮏࡢ⮫ᗋศ㞳ᰴ 25 ᰴࡢẚ㍑ࢤࣀ࣒ ゎᯒࡢᨭࠋ ᨭࡣᖹᡂ 23 ᖺᗘࡽ 27 ᖺᗘࡲ࡛ࠊ ᭱ึ ShT-P ࠾ࡼࡧ ShT-N ࡢ 2 ᰴࡘ࠸࡚ Roche454GS FLX ࢆ⏝࠸ࡓゎᯒࠋḟ ShT-P ᰴ㛵ࡋ࡚ࣉࣛࢫ࣑ࢻࢆ 㝖ࡃ⣙ 250 ᡤࡢࢠࣕࢵࣉࢡ࣮ࣟࢪࣥࢢࠋࡉࡽ Illumina ࢆ⏝࠸ࡓࢩ࣮ࢡ࢚ࣥࢫࡢ☜ㄆࠋḟࠊ᪥ᮏ ࡢ⮫ᗋศ㞳ᰴ 25 ᰴࡘ࠸࡚ࡢ Illumina ࢆ⏝࠸ࡓࢩ ࣮ࢡ࢚ࣥࢫゎᯒࠋࡉࡽࣉࣛࢫ࣑ࢻゎᯒࢩ࣮ࢡ࢚ ࣥࢫࡢ☜ㄆࢆ┠ⓗࡋ࡚ࠊShT-P ᰴ㛵ࡋ࡚ PacBio ࢆ⏝࠸ࡓゎᯒࠋࡑࢀࡽຍ࠼࡚ࠊ┤᥋ࡢᢏ⾡ᣦᑟ ࡋ࡚ࠊࢤࣀ࣒ࢹ࣮ࢱࡢࣀࢸ࣮ࢩࣙࣥἲ࠾ࡼࡧゎ ᯒ᪉ἲࡘ࠸࡚ࡢᣦᑟࢆࠊࡲࡓ㑇ఏ◊࡚ࢫ࣮ࣃ࣮ ࢥࣥࣆ࣮ࣗࢱ࣮⏝ࡢᣦᑟࢆཷࡅࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᡃ ࠎ ࡣ ᪥ ᮏ ࡛ ᭱ ึ ศ 㞳 ࡉ ࢀ ࡓ ”Mycobacterium ulcerans subsp. shinshuense” ATCC 33728 ᰴࡘ ࠸࡚(ẘᛶ⬡㉁ mycolactone ࢆ⏘⏕ࡋ㯤Ⰽࢥࣟࢽ࣮ࠊ ShT-P ᰴ㸧Roche 454 GS FLX ࢆ⏝ࡋ࡚ ࢤࣀ࣒ ゎᯒࢆヨࡳࠊ⣙ 250 ᡤࡢ gap ࢆ close ࡍࡿࡇ࡛ ࢡࣟࣔࢰ࣮࣒ࡢࢩ࣮ࢡ࢚ࣥࢫࢆ⋓ᚓࡋࠊࡉࡽ PacBio ࡛ࡢゎᯒ⤖ᯝࢆຍ࠼ࠊࡼࡾ⢭ᗘࡢ㧗࠸ࢩ࣮ ࢡ࢚ࣥࢫ⤖ᯝࢆᚓࡓࠋM. shinshuense ATCC33728 ᰴ(5,899,681 bps)ࡣつᶍ࡞ re-arrangement ࡀ ⏕ࡌ࡚࠸ࡿࡇࡀ☜ㄆࡉࢀࠊ5027 ಶࡢ CDS, 46 ಶࡢ tRNA, 451 ಶࡢ pseudogene, 245 ಶࡢ PE/PPE family (࠺ࡕ 96 ಶࡣ pseudogene), 221 ಶࡢ insertion sequence (࠺ࡕ 37 ಶࡣḞᦆኚ␗)ࢆ᭷ࡍࡿࡇࡀุ ᫂ࡋ࡚࠸ࡿࠋࡇࡢ⤖ᯝࡣࠊM. marinum ࡢ⣙ 6.6Mbps ẚ㍑ࡍࡿᑠࡉ࠸ࡶࡢࡢࠊ࣮࢞ࢼ࡛ศ㞳ࡉࢀࡓ M. ulcerans Agy99 ᰴࡢ 5.6Mbps ࡼࡾࡁ࠸ࠋࡲࡓࠊ CDS ᩘࡣ M. marinum ࡀ 5424 ಶࠊAgy99 ࡀ 4160 ಶࠊM. shinshuense5027 ಶ࡛ࠊࡉࡽ pseudogene ᩘࡣࠊM. marinum ࡀ 65 ಶᑐࡋ࡚ࠊAgy99 ࡣ 771 ಶࠊM. shinshuense ࡀ 245 ಶ࡛ࠊM. shinshuense ࡣ M. marinum Agy99 ࡢ୰㛫ᆺࢆ♧ࡋ࡚࠸ࡿࡶࡢ ⪃࠼ࡽࢀࡓࠋ⌧ᅾࠊ᪥ᮏࡢ⮫ᗋศ㞳 25 ᰴ(᭱ึࡢ ᨭࢆཷࡅࡓ ATCC37728 ࢆྜࢃࡏ࡚ 26 ᰴ)ࠊ ࡢ M. ulcerans ࡢ┦㐪➼ࢆゎᯒ୰࡛࠶ࡿࠋ ࡋࡋ࡞ࡀࡽࠊ ཎᛶࡢせ┠ࡉࢀࡿᕧ plasmid ࡣࠊከࡃࡢ⧞ࡾ㏉ࡋ㓄ิ㧗࠸ GC ྵ㔞ࡢࡓࡵࠊ PacBio ࡛ࡢゎᯒ࡛ࡶ㝈⏺ࡀ࠶ࡿࡶࡢᛮࢃࢀࠊṇ☜ ࡞ࢩ࣮ࢡ࢚ࣥࢫゎᯒࡲ࡛ࠊ୍Ṍࡢ≧ែ࡛࠶ࡿࠋࡲ ࡓࠊ୍ࡘࡢ⳦ࡘ࠸࡚ࠊplasmid ࡀ 1-2 ࢥࣆ࣮ࡢࡳ ࡞ࡢ࡛ࠊplasmid ࡢศ㞳ࡶ㞴⯟ࡋ࡚࠾ࡾࠊplasmid ࡢゎᯒࡀḟࡢㄢ㢟࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏ㑥ࡢࣈ࣮ࣝࣜ₽⒆ᝈ⪅ࡽศ㞳ࡉࢀࡓ⮫ᗋศ㞳 26 ᰴࡢゎᯒ࡛ࠊࡇࢀࡲ࡛ࢧ࣮ࣥ࢞ἲࡼࡿ୍㒊ࡢ▷࠸ ࢩ࣮ࢡ࢚ࣥࢫゎᯒࡔࡅ࡛ࡣᅔ㞴࡛࠶ࡗࡓࠊศ㞳ᰴ㛫 ࡢ␗ྠᛶࡢゎᯒࡀ㐍ᒎࡋࡘࡘ࠶ࡿࠋࡇࢀࡼࡗ࡚᪥ ᮏᅜෆࡢศᏊᏛⓗ▱ぢࡀ✚ࡉࢀࡿࡶࠊࣈ ࣮ࣝࣜ₽⒆ᑐ⟇ࡢᛂ⏝ࡀྍ⬟࡞ࢁ࠺ࠋ ᪉ ࠊ ࡇ ࢀ ࡲ ࡛ ࠊ M. ulcerans ྠ ᵝ ࡢ mycolactone ᵝࡢẘᛶ⬡㉁ࢆ code ࡍࡿᕧ plasmid ࢆ ᭷ ࡍ ࡿ ᢠ 㓟 ⳦ ⩌ (mycolactone producing mycobacteria㸸MPM)ࡀ⨯ᝈࡋࡓ࢚࢝ࣝࡸ㨶ࡽศ 㞳ࡉࢀ࡚࠸ࡿࠋᮏ㑥࡛ࡶࠊ⨯ᝈࡋࡓ㨶⩌ࡸ࣓࢝࡞ ࡽ M. ulcerans ㏆⦕ࡢᢠ㓟⳦ࡀศ㞳ࡉࢀ࡚࠾ࡾࠊ ࡇࢀࡽ᪥ᮏࡢ MPM ◊✲ࡢⓎᒎࡶ⪃࠼ࡽࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Yotsu, RR. et al.,J. 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Dermatol. 42,588-595(2015) - 451 - බເᨭㄢ㢟㸸2012ࠥ2013 ᖺᗘ 㧗ཎᛶࢫࢺࣞࣉࢺࢥࢵ࢝ࢫ࣭࣑ࢳࢫࡣᏑᅾࡍࡿ㸸ࡑࡢཎᛶᏛࡢ◊✲ ᇶ┙◊✲㸦㹁㸧 㛗᐀ ⚽࣭᫂ᚨᓥᏛᏛ㝔ࢯࢩ࢜ࢸࢡࣀࢧ࢚ࣥࢫ◊✲㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ࣄࢺཱྀ⭍ෆᖖᅾᛶࡢ㐃㙐⌫⳦ࡢ୍✀࡛࠶ࡿ Streptococcus mitis㸦SM㸧ࡣࠊ୍⯡ⓗ㠀࣭పཎ ᛶࡢ᪥ぢឤᰁ⳦ࡋ࡚ㄆ㆑ࡉࢀ࡚࠸ࡿࠋࡋࡋ࡞ ࡀࡽࠊSM ࡣ௦⾲ⓗ࡞ࣄࢺཎᛶࡢ㐃㙐⌫⳦࡛࠶ࡿ⫵ ⅖⌫⳦㸦S. pneumoniae㸧ࡢ㢮⦕⳦࡛࠶ࡿࡇຍ ࠼ࠊࡇࢀࡽࡢ⳦✀࡛ᵓᡂࡉࢀࡿ࣑ࢳࢫ-⫵⅖⌫⳦⩌㐃 㙐⌫⳦㸦MGS㸧ࡢࡢᵓᡂ⳦✀࠾࠸࡚ࡶࠊࣄࢺ ᑐࡍࡿཎᛶࡢ㛵㐃㛵ࡍࡿሗ࿌ࡀ㏆ᖺᩓぢࡉࢀ ࡿࡼ࠺࡞ࡾࠊMGS ࡢࣄࢺࡢཎᛶࡀᨵࡵ࡚ὀ┠ ࡉࢀ࡚࠸ࡿࠋᡃࠎࡣࠊඛ⾜◊✲࠾࠸࡚ SM ࡢ୰ ⭷Ꮝᙧᡂẘ⣲࡛࠶ࡿࢥࣞࢫࢸ࣮ࣟࣝ౫Ꮡᛶ⣽⬊⁐ゎ ẘ⣲㸦CDC㸧ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࢆಖ᭷ࡋࠊᙉ࠸⣽ ⬊㞀ᐖᛶࢆ♧ࡍࡢࡳ࡞ࡽࡎࠊș-ࣛࢡࢱ࣒⣔ᢠ⏕ ⪏ᛶࢆ♧ࡍ SM ࡀᏑᅾࡍࡿࡇࢆぢฟࡋࡓࠋ ຍ࠼࡚ࠊ ࡇࢀࡲ࡛ࠊSM ࠾࠸࡚⫵⅖⌫⳦ࡢࢤࣀ࣒㞧ࢆ ♧၀ࡍࡿሗ࿌ࡶᡂࡉࢀ࡚࠸ࡿࠋࡑࡇ࡛ᡃࠎࡣࠊࡇࡢ ࡼ࠺࡞ SM ࡢㅖ≉ᛶࢆࢤࣀ࣒ሗᇶ࡙࠸࡚᫂ࡽ ࡋ࡚࠸ࡃࡇࢆ┠ⓗࡋ࡚ࠊࢤࣀ࣒ᨭᴗࡢࢧ ࣏࣮ࢺࢆཷࡅ࡚ SM ᰴࡢࢤࣀ࣒㓄ิࡢྲྀᚓゎᯒࢆ ⾜ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 2012 ᖺᗘࡣࠊ9 ᰴࡢ SM ᰴ 1 ᰴࡢ SM 㢮⦕ᰴࢆ ᑐ㇟ࡋࠊillumina MiSeq ࠾ࡼࡧ Roche 454 GS FLX ࢆ⏝࠸࡚㓄ิゎᯒࢆ⾜࠸ࠊNewbler ࡼࡿࢭ ࣥࣈࣜࢆ⾜ࡗࡓࠋ2013 ᖺᗘࡣࠊࡉࡽ 36 ᰴࡢ SM ᰴ 4 ᰴࡢ SM 㢮⦕ᰴࡢィ 40 ᰴࡘ࠸࡚ࠊillumina MiSeq ࢆ⏝࠸ࡓࢻࣛࣇࢺࢤࣀ࣒ࡢ㏣ຍྲྀᚓࢆ⾜ࡗࡓࠋ ࡉࡽ 2014 ᖺᗘࡣࠊඹྠ◊✲ࡋ࡚ࠊCDC 㑇ఏ Ꮚࡢಖ᭷ࣃࢱ࣮ࣥࢆᣦᶆ௦⾲ⓗ࡞ SM ᰴࢆ 10 ᰴ㑅 ᐃࡋ࡚ࢤࣀ࣒ࣇࢽࢵࢩࣥࢢࢆヨࡳࡿඹࠊࡑࡢ ෆࡢ 2 ᰴࡘ࠸࡚ࡣ PacBio ࢆ⏝࠸ࡓゎᯒࡶ⾜ࡗࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡲࡎࠊ2012 ᖺᗘࡢゎᯒ࡛ࡣࠊSM ᰴ࠾ࡼࡧ SM 㢮 ⦕ᰴ㸦ィ 10 ᰴ㸧ࡢࢻࣛࣇࢺࢤࣀ࣒ሗ㸦coverage㸸 59.9ࠥ152.1ࠊcontig ᩘ㸸30ࠥ68ࠊscaffold ᩘ㸸 2ࠥ5㸧ࢆᚓࡓࠋᚓࡽࢀࡓࢤࣀ࣒㓄ิሗᇶ࡙࠸࡚ Average nucleotide index㸦ANI㸧ゎᯒࡼࡿࢤࣀ࣒ 㓄ิࣞ࣋ࣝࡢẚ㍑᳨ウࢆ⾜ࡗࡓ⤖ᯝࠊSM ࡣள⣔⤫ ศ㢮ࡍࡿࡢࡀ㐺ᙜุ᩿ࡉࢀࡿࡼ࠺࡞⣔⤫ࡢ␗࡞ ࡿ⳦ᰴ⩌ࡢᏑᅾࡀᙉࡃ♧၀ࡉࢀࡓࠋࡇࡢ⤖ᯝࡼࡾࠊ SM ࡣࢤࣀ࣒ᵓ㐀ࡢከᵝᛶࡀⴭࡋࡃࠊSM ࢆ⣔⤫Ꮫⓗ ᳨ウࡍࡿࡓࡵࡣࡉࡽ SM ᰴࡢࢤࣀ࣒ሗࡀᚲ せ࡛࠶ࡿ࠸࠺⤖ㄽ⮳ࡗࡓࠋ ࡇࢁ࡛ࠊ⫵⅖⌫⳦ྠࡌࡃࠊSM ࠾࠸࡚ࡶ⸆ ⪏ᛶ⳦ࡢⓎ⏕ࡀᠱ㡯࡞ࡗ࡚࠸ࡿࠋ⫵⅖⌫⳦ ࠾ࡅࡿș-ࣛࢡࢱ࣒⣔ᢠ⏕ᑐࡍࡿ⪏ᛶࡣࠊ࣌ࢽ ࢩࣜࣥ⤖ྜࢱࣥࣃࢡ㉁㸦PBP㸧࠾ࡅࡿ≉ᐃࡢ࣑ ࣀ㓟ኚ␗㉳ᅉࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋࡑࡇ࡛ ᚓࡽࢀࡓ SM ᰴࡢࢻࣛࣇࢺࢤࣀ࣒㓄ิሗࡽ PBP ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࡢ㓄ิሗࢆᢳฟࡋ᳨࡚ウࡋࡓ ⤖ᯝࠊș-ࣛࢡࢱ࣒⣔ᢠ⏕⪏ᛶᐤࡍࡿ⪃࠼ ࡽࢀ࡚࠸ࡿ PBP ࡢ࣑ࣀ㓟㓄ิኚ␗ࢆక࠺㑇ఏᏊ㓄 ิࡢኚ␗ࡀ☜ㄆࡉࢀࠊSM ࡢș-ࣛࢡࢱ࣒⣔ᢠ⏕≀㉁ ⪏ᛶࡣ 3 ✀ࡢ PBP㸦PBP1AࠊPBP2BࠊPBP2X㸧 ࠾ࡅࡿ⪏ᛶ࣑ࣀ㓟Ⅼኚ␗ࡀ┦ຍⓗᐤࡋ࡚ ࠸ࡿࡇࡀ᥎ ࡉࢀࡓࠋ ⥆ࡃ 2013 ᖺᗘࡢゎᯒ࡛ࡣࠊࡉࡽ 36 ᰴࡢ SM ࡘ࠸࡚ࢻࣛࣇࢺࢤࣀ࣒ሗ㸦coverage㸸73.1ࠥ 209.4ࠊscaffold ᩘ㸸25ࠥ119㸧ࢆᚓࡓࠋᚓࡽࢀࡓࢤ ࣀ࣒㓄ิሗࡼࡾ⫵⅖⌫⳦ࡢ MLST ゎᯒ࡛⏝ࡉࢀ ࡚࠸ࡿ 4 ✀ࡢࣁ࢘ࢫ࣮࢟ࣆࣥࢢ㑇ఏᏊ㸦 ddl, gdh, rpoB, sodA㸧ࡢ㒊ศ㓄ิࢆᢳฟࡋ࡚ࢥࣥ࢝ࢸ࣐࣮ ࡋࠊClustalW ࡛࣓ࣛࣥࣥࢺᚋ MEGA ࢆ⏝࠸ ࡚⣔⤫ᶞࢆసᡂࡋࡓࠋࡑࡢ⤖ᯝࠊSM ࡢᇶ‽ᰴࢆྵࡴ ࢿ࢜ࢱࣉ⩌࠾ࡼࡧ⫵⅖⌫⳦⩌ࠊࡉࡽ SM ⫵⅖ ⌫ ⳦ ࡢ ࣁ ࣈ ࣜ ࢵ ࢻ ᰴ ⓗ ࡞ ≉ ᛶ ࢆ ♧ ࡍ S. pseudopneumoniae㸦SPpn㸧ࡣࡑࢀࡒࢀࢡ࣮ࣞࢻࢆ ᙧᡂࡋࡓࡀࠊࡢ SM ᰴ࡛ࡣ≉ᐃࡢࢡ࣮ࣞࢻࢆᙧᡂ ࡋ࡞ࡗࡓࠋ ࡉࡽ 2014 ᖺᗘࡣࠊCDC ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊ ࡢಖ᭷ࣃࢱ࣮ࣥࢆᣦᶆࡋ࡚௦⾲ⓗ࡞ SM ᰴࢆ 10 ᰴ 㑅ᐃࡋࠊࢤࣀ࣒ࣇࢽࢵࢩࣥࢢࢆヨࡳࡓࠋࡋࡋ࡞ ࡀࡽࠊ20bp ⛬ᗘࡽᩘⓒ bp ࡢࢱࣥࢹ࣒ࣜࣆ࣮ࢺ ࡽᡂࡿࢠࣕࢵࣉࡢᏑᅾࡼࡾࠊࢧ࣮ࣥ࢞ἲࡼࡿࣇ ࢽࢵࢩࣥࢢࡣᅔ㞴࡛࠶ࡗࡓࠋࡑࡇ࡛ࠊ10 ᰴ୰ࡢ 2 ᰴࡘ࠸࡚ PacBio ࡛ࡢゎᯒࢆ⾜ࡗࡓ⤖ᯝࠊ2 ᰴ୰ࡢ 1 ᰴࡘ࠸࡚㛗㓄ิࡢྲྀᚓᡂຌࡋࡓࠋ࡞࠾ࠊ ࡢᰴࡘ࠸࡚ࡣ⤖ᯝⓗࣇࢽࢵࢩࣥࢢࢆ㐩ᡂ࡛ࡁ ࡞ࡗࡓࡀࠊࡑࢀࡒࢀࡢᰴࡢࢤࣀ࣒ෆᏑᅾࡍࡿࢱ ࣥࢹ࣒ࣜࣆ࣮ࢺࡢከࡃࡘ࠸࡚ࠊࡑࡢᵓ㐀ࢆỴᐃࡍ ࡿࡇࡀ࡛ࡁࡓࠋ ࡇࢁ࡛ࠊSM ࡛ࡣࠊ≉ᚩࡀ␗࡞ࡿ 3 ✀ࡢ CDC㹙ձ ⫵⅖⌫⳦ࡀ⏘⏕ࡍࡿ CDC ࡛࠶ࡿ pneumolysin ࡢ࣍ ࣔࣟࢢ࡛࠶ࡿ mitilysin㸦MLY㸧ࠊղ SM ࡢ Nm-65 ᰴ ࡀ ⏘ ⏕ ࡍ ࡿ SM ⏤ ᮶ ࣄ ࢺ ⾑ ᑠ ᯈ จ 㞟 ᅉ Ꮚ 㸦Sm-hPAF㸧ࠊճ ᡃࠎࡀ᪂ࡓぢฟࡋࡓ 5 ࢻ࣓ࣥ ᆺ CDC㸦ExD-CDC㸧㹛ࢆᵝࠎ࡞ࣃࢱ࣮࡛ࣥ」ᩘಖ᭷ ࡍࡿᰴࡢᏑᅾࡀ☜ㄆࡉࢀ࡚࠸ࡿࠋᅇࡢゎᯒ࠾ࡅ ࡿ⿕᳨ᰴࡘ࠸࡚ CDC 㑇ఏᏊࡢಖ᭷ࣃࢱ࣮ࣥᇶ ࡙࠸ࡓ᳨ウࢆ⾜ࡗࡓ⤖ᯝࠊCDC 㑇ఏᏊࡢಖ᭷ࣃࢱ࣮ ࣥࡘ࠸࡚ࡣ⣔⤫ⓗ᫂☜࡞つ๎ᛶࡣ☜ㄆࡉࢀ࡞ ࡗࡓࠋࡋࡋ࡞ࡀࡽࠊྛ CDC 㑇ఏᏊࡢ࿘㎶㓄ิࣇ ࢛࣮࢝ࢫࡋࡓゎᯒࡢ⤖ᯝࡽࠊSm-hPAF ࠾ࡼࡧࡑࡢ ࣍ࣔࣟࢢࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࡢ࿘㎶ᵓ㐀ࡢಖᏑᛶࡣ 㧗ࡃࠊ≉ୖὶᇦࡣࡰಖᏑࡉࢀ࡚࠸ࡿࡇ ࡀุ᫂ࡋࡓࠋࡲࡓࠊୗὶᇦࡢ㑇ఏᏊᵓ㐀ࡣࡁࡃ 2 ࣃ ࢱ ࣮ ࣥ ศ 㢮 ࡉ ࢀ ࠊ ࡑ ࡢ ୍ ᪉ ࡣ SPpn ⏤ ᮶ Sm-hPAF ࣍ࣔࣟࢢࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࡢୗὶᇦ ྠᵝࡢ㑇ఏᏊᵓ㐀࡛࠶ࡗࡓࠋࡲࡓࠊSM ࢜ࣜࢪࢼࣝࡢ ExD-CDC ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࡢ࿘㎶ᵓ㐀ࡘ࠸࡚ ࡶẚ㍑ⓗಖᏑࡉࢀ࡚࠾ࡾࠊSm-hPAF ྠᵝୖὶᇦ - 452 - ࡢ㑇ఏᏊᵓ㐀ࡢಖᏑᛶࡀ㧗ࡃࠊୗὶᇦࡣࡁࡃ 2 ࣃ ࢱ࣮ࣥࡢ㑇ఏᏊᵓ㐀ศ㢮ࡉࢀࡓࠋ୍᪉ࠊMLY ࢆࢥ ࣮ࢻࡍࡿ㑇ఏᏊࡢ࿘㎶ᵓ㐀ࡣẚ㍑ⓗከᵝᛶࡀ࠶ࡿ ࡇࡀ☜ㄆࡉࢀࡿඹࠊࡇࡢ㡿ᇦࡀ scaffold ࡢᮎ ➃࡞ࡗ࡚࠸ࡿゎᯒ⤖ᯝࡶከࡃぢࡽࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢤࣀ࣒ᨭᴗࡼࡾᚓࡽࢀࡓ SM ࡢࢤࣀ࣒㓄 ิࢆ⏝࠸ࡓ㧗⢭ᗘ࡞⣔⤫ゎᯒࡼࡾࠊከᵝ࡞ SM ᰴ ࡽᡂࡿ SM ࡢ㞟ᅋᵓ㐀ࡀ᫂ࡽ࡞ࡾࠊࡉࡽ CDC ࣇ࣑࣮ࣜࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊ㡿ᇦࡢゎᯒ ࡼࡗ࡚ࠊSM ࡀಖ᭷ࡍࡿࡇࢀࡽࡢ㑇ఏᏊࡢ࿘㎶ᵓ㐀ࡣ CDC ࡢ✀㢮ࡈࣃࢱ࣮ࣥࡀྍ⬟࡛࠶ࡗࡓࠋࡑࢀ ࡽࡢ⤖ᯝࡽࠊSM ᰴࡢ㐍ᑐᛂࡋࡓ CDC ࢆࢥ࣮ ࢻࡍࡿ㑇ఏᏊࡢࣞࣃ࣮ࢺ࣮ࣜࡢኚ࠾ࡼࡧከᵝᛶࡢ ᣑࡀ᥎ᐃࡉࢀࡓࠋࢤࣀ࣒ᨭᴗᇶ࡙࠸ࡓࡇࢀ ࡽࡢ◊✲ᡂᯝࡽࠊࣄࢺᑐࡋ࡚ཎᛶࢆ♧ࡍ⫵⅖ ⌫⳦ࡢ㢮⦕⳦࡛࠶ࡾ࡞ࡀࡽᮍࡔ᫂࡞Ⅼࡀከ࠸ SM ࡘ࠸࡚ࠊCDC ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࢆಖ᭷ࡋ࡚ ⣽⬊㞀ᐖᛶࢆⓎࡍࡿࢧࣈࢢ࣮ࣝࣉࡢཎᛶࡢホ ౯㈨ࡍࡿ㔜せ࡞ሗࢆᥦ౪࡛ࡁࡿࡶࡢᮇᚅࡉࢀ ࡿࠋ ᚋࡢᒎᮃࡋ࡚ࠊSM ࠾ࡼࡧࡑࡢ㢮⦕ᰴࢆྵࡵࡓ ࢤࣀ࣒㓄ิࡢሗࢆ⏝࠸ࡓ㧗⢭ᗘ࡞⣔⤫ゎᯒࢆࡉࡽ 㐍ࡵࡿࡇࡼࡾࠊMGS ࡢ⣔⤫㐍ⓗ࡞⨨ࡅ ࡸ SM ࡢ㧗ཎᛶᰴࡢฟ⌧࣓࢝ࢽࢬ࣒ࡢゎ᫂⧅ࡀ ࡿሗࡀᚓࡽࢀࡿࡢ࡛ࡣ࡞࠸ᮇᚅࡋ࡚࠸ࡿࠋࡑ ࡋ࡚ᮏ◊✲ࢆࠊࣄࢺཎయࡋ࡚㔜せ࡞⫵⅖⌫⳦ࢆ ྵࡴ MGS ࡼࡿឤᰁண㜵ࡸᝈ࣭ែࡢ⒪ࠊ ࡲࡓ SM ࡢ⸆⪏ᛶࡢᢚไ࡞㈉⊩ࡍࡿ᭷ព⩏ ࡞ሗࢆᥦ౪࡛ࡁࡿ㔜せ࡞ឤᰁࢤࣀ࣒་Ꮫ◊✲ ࡋ࡚ࠊࡉࡽⓎᒎࡉࡏ࡚࠸ࡁࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ㄽᩥ‽ഛ୰ - 453 - බເᨭㄢ㢟㸸2010 ᖺᗘ ṑ࿘⣽⳦ࡢཎࣉࣟࢸ࣮ࢮ࣭ࢻ࣊ࢪࣥࡢ⭷㍺㏦࣭ศἪᶵᵓࡢᐜゎ᫂ ᇶ┙◊✲㸦㸿㸧 ୰ᒣ ᾈḟ࣭㛗ᓮᏛᏛ㝔་ṑ⸆Ꮫ⥲ྜ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ṑ࿘⅖ࡣ ṓ௨ୖࡢ᪥ᮏேࡢ༙ᩘ௨ୖࡀ⨯ᝈࡋ࡚࠸ ࡿ៏ᛶឤᰁ࡛࠶ࡾࠊṑࡢ႙ኻࡢ᭱ࡢཎᅉ࡛࠶ࡿࠋ ᡃࠎࡣࡇࢀࡲ࡛೫ᛶ᎘Ẽᛶ⣽⳦࡛࠶ࡿṑ࿘ཎ⳦ ⩌࠾ࡅࡿࣉࣟࢸ࣮ࢮ࣭ࢻ࣊ࢪࣥࡢ⭷㍺㏦ࠊ ཎᅉᏊ㍺㏦ࡢᶵᵓࡢゎᯒࢆ⾜࠸ࠊ᪂ࡓ࡞ศἪ㍺㏦⣔ࠊ 7\SH,; ศἪᶵᵓࢆ 3RUSK\URPRQDVJLQJLYDOLV ࡽ ぢฟࡓࠋࡑࡢᶵ⬟ࡘ࠸࡚ 3JLQJLYDOL ࠾࠸࡚ኚ ␗ᰴࢆ⏝࠸ࡓヲ⣽࡞ゎᯒࢆ⾜ࡗ࡚ࡁࡓࠋṑ࿘ཎ⳦ ⩌ࡢ୍ࡘ࡛࠶ࡿ 3UHYRWHOODLQWHUPHGLD ࡶᮏᶵᵓ ࡢ㢮ఝࡢ㑇ఏᏊࡀศᕸࡍࡿࡇࢆ☜ㄆࡋ࡚࠸ࡿࠋࡋ ࡋᮏ⳦࡛ࡣ㑇ఏᏊ᧯స࡞ࡢศᏊ⏕≀ⓗ◊✲ࡢᇶ ┙ࡀ☜❧ࡋ࡚࡞࠸Ⅽゎᯒࡀ㐍ࢇ࡛࠸࡞࠸ࠋࡑࡇ࡛ 3LQWHUPHGLD࠾࠸࡚ࡢཎᅉᏊ㍺㏦ᶵᵓࡢ⥙⨶ ⓗ࡞ゎᯒࢆ┠ᣦࡋࡓࠋࡇࢀࡲ࡛ணഛⓗᐇ㦂ࡼࡾ 3LQWHUPHGLD ࡢ⮫ᗋศ㞳ᰴ ᰴ20$ࡽḟୡ௦ ࢩ࣮ࢡ࢚ࣥࢧ࣮㸦5RFKH㸧ࢆ⏝࠸ࡓゎᯒࡼࡾࠊ ಶࡢ FRQWLJ ࡽ࡞ࡿ GUDIW 㓄ิࢆᚓ࡚࠸ࡓࠋࡑࡇ ࡛ᮏ◊✲ᨭࡼࡾࡇࡢ 3LQWHUPHGLDࡢ⮫ᗋศ㞳 ᰴ ᰴࡢࢤࣀ࣒㓄ิࢆỴᐃࡋࠊࡑࡢࢤࣀ࣒ᵓ㐀ࡢ ゎᯒࠊࢤࣀ࣒ሗࡢබ㛤ࢆ┠ᣦࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࡇࢀࡲ࡛ 3LQWHUPHGLD ࡢ⮫ᗋศ㞳ᰴ ᰴ20$ ࡽḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮㸦5RFKH㸧ࢆ⏝࠸ࡓゎ ᯒࡼࡾࠊ ಶࡢ FRQWLJ ࡽ࡞ࡿ GUDIW 㓄ิࢆᚓ࡚ ࠸ࡓࡀࠊNE ㉸ࡢ㔜」㓄ิࡢᏑᅾࡢⅭࡇࢀ௨ୖࡢゎ ᯒࢆ㐍ࡵࡿࡇࡀ࡛ࡁ࡞ࡗࡓࠋࡑࡇ࡛ᮏࢤࣀ࣒ᨭ ࢆά⏝ࡍࡿࡇࡼࡾࠊ 3UHLQWHUPHGLD 20$ ᰴࡢ㛗ࡢࢤࣀ࣒㓄ิࢆỴᐃࡋࠊࡑࡢヲ⣽࡞ゎ ᯒࢆ┠ᣦࡋࡓࠋ ලయⓗࡣୗグࡢᨭࢆ౫㢗ࡋࡓࠋ 3UHLQWHUPHGLD20$ ᰴࡢ IRVPLGࣛࣈࣛࣜ ࣮ࡢసᡂࠋ ୖグࣛࣈ࣮ࣛࣜࡽࣛࣥࢲ࣒㑅ᢥࡋࡓ⣙ ࢡ࣮ࣟࣥ㸦㔜」ᗘ㸹㹶㸧ࡢ୧ᮎ➃㓄ิࡢỴ ᐃࠋ ୖグࡢ⤖ᯝࢆ⏝ࡋࡓ FRQWLJ ࡢᩚิࠊ࠾ࡼࡧᩚ ิࡋࡓ FRQWLJ 㛫ࡢ JDSFORVLQJ㸦3&5 ࡼࡿ FORVLQJ IRVPLGFORQH ࡢሷᇶ㓄ิỴᐃ㸧ࠋ 3UHLQWHUPHGLD20$ ᰴࡢࢤࣀ࣒ሷᇶ㓄ิࡢ ࢡ࢜ࣜࢸ࣮ࢆୖࡆࡿࡓࡵࡢ ,OOXPLQD ࠶ࡿ࠸ࡣ 62/LG ࢆ⏝࠸ࡓ UHVHTXHQFLQJ ゎᯒࠋࡇࢀࡲ࡛ࡢ GDWD㸦ḟୡ௦ࢩ࣮ࢡ࢚ࣥࢧ࣮㸦5RFKH㸧ゎᯒࠊ IRVPLG ࣛ ࣈ ࣛ ࣜ ࣮ ゎ ᯒ 㸧 ྜ ࢃ ࡏ ࡚ DVVHPEOH ࢆ࠾ࡇ࡞࠺ࠋ 㸦㸲㸧࡛ࡢゎᯒ⤖ᯝࡢ FRQWLJ ᩚิࡀṇࡋ࠸☜ ㄆࡍࡿࠋ࣓ࢱࣀ࣮ࣝᅛᐃࡋࡓ⳦యࢆ⏝࠸࡚≀⌮ ⓗ࡞ไ㝈㓝⣲ษ᩿ࣃࢱ࣮ࣥࢆ RSWLFDOPDSSLQJ ἲ᳨࡚ฟࠊ㸦㸲㸧ࡢࢤࣀ࣒㓄ิࡽண ࡉࢀ ࡿไ㝈㓝⣲ษ᩿ࣃࢱ࣮ࣥẚ㍑ࠊ᳨ドࡍࡿࠋ ᳨ド῭ࡳࡢ GDWD ࡽ㑇ఏᏊᢳฟࢆ 0L*$3 ࢯࣇࢺ ࡚⾜࠺ࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᅇࡢᨭࡼࡾ KLJKTXDOLW\ ࡞ 3UHLQWHUPHGLD 20$ ᰴࡢ㛗ࢤࣀ࣒㓄ิࢆỴᐃࡋࡓࠋᮏ⳦ࡣ ಶࡢ⎔≧ᰁⰍయࠊES ES ࢆᣢࡕࠊ ࡑࢀࡒࢀ ಶ ಶࡢ㑇ఏᏊࡀᏑᅾࡍࡿࡇ ࢆ᫂ࡽࡋࡓࠋ3UHLQWHUPHGLD ࢤࣀ࣒㓄ิࡘ ࠸࡚ࡣࡇࢀࡲ࡛㸯ᰴ㸦VWUDLQ㸧࡛ሗ࿌ࡉࢀ࡚࠸ ࡿࡀࠊࡑࡢᰴࡢࢤࣀ࣒ẚ㍑ࡍࡿࠊ➨ࢡࣟࣔࢰ ࣮࣒ࡀ⣙ NE ࡁ࠸ࡇࠊつᶍ࡞ࢤࣀ࣒ᵓ㐀ࡢ ⦅ᡂࡀࡳࡽࢀࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ᚓࡽࢀࡓࢤࣀ࣒㓄ิࡣ 7\SH,; ศἪᶵᵓᚲ㡲ࡢ 㑇ఏᏊ⩌ࡀ࡚Ꮡᅾࡍࡿࢆ☜ㄆࡋࡓࠋ ࡲ ࡓ ࠊ ࢤ ࣀ ࣒ 㓄 ิ ࡢ ࡉ ࡽ ࡞ ࡿ ゎ ᯒ ࡼ ࡾ ࠊ 3UH LQWHUPHGLD ࡢࢤࣀ࣒ࡣከᵝከᩘࡢ㌿⛣ᛶ㑇ఏᅉ ᏊࡀᏑᅾࡍࡿࡇࢆ᫂ࡽࡋࡓࠋ20$ ᰴࡣ FRQMXJDWLYH WUDQVSRVR &7Q ࡀ ✀ ࢥ ࣆ ࣮ PRELOL]DEOHWUDQVSRVRQ07Qࡀ ✀ ࢥࣆ࣮Ꮡᅾ ࡋ࡚࠸ࡓࠋࡲࡓVWUDLQ ࡢࢤࣀ࣒ࡣ &7㹬 ✀ ࢥࣆ࣮ࠊ07㹬 ✀ ࢥࣆ࣮ࡀᏑᅾࡋ࡚࠸ࡓࠋ๓㏙ࡢࢤ ࣀ࣒ࢧࢬࡢᰴ㛫ࡢᕪࡣࡇࢀࡽࡢ」ᩘࡢ㌿⛣ᛶ㑇ఏ ᅉᏊࡢᤄධࡼࡿࡇࡀ♧၀ࡉࢀࡓࠋࡇࡢࡼ࠺ 20$ ᰴࡢࢤࣀ࣒ࡣᩘ༑ NES ࡢ &7Q ࡀ」ᩘᏑᅾࡍࡿ ࡽ )RVPLG ࢡ࣮ࣟࣥࢆ⏝࠸ࡓゎᯒࠊRSWLFDO PDSSLQQJ ࡼࡿ㓄ิ☜ㄆ࡞ࡋࡣࢤࣀ࣒㓄ิࢆỴ ᐃࡍࡿࡇࡣྍ⬟࡛࠶ࡗࡓࡇࡶ♧ࡉࢀࡓࠋ ࡲࡓࢤࣀ࣒㛗ࢃࡓࡗ࡚ ES ࡢ㓄ิࡀ 20$ ᰴࡽ ಶࠊVWUDLQ ᰴࡽ ಶぢ࠸ࡔ ࡉࢀࡓࠋKRPRORJ\ ᳨⣴≉␗ⓗ࡞ 3&5 ࡼࡿゎᯒ ࡽ 3UHLQWHUPHGLD 3UHQLJUHVFHQV ➼ࡢ㏆⦕✀ ௨እࡣࡇࡢ㓄ิࡣ᳨ฟࡉࢀ࡞ࡗࡓࠋࡇࢀࡽࡢ⤖ ᯝ ࡽ ࡇ ࡢ 㓄 ิ ࡀ 3UH LQWHUPHGLD 3UH QLJUHVFHQV ≉␗ⓗ࡞ࡶࡢ⪃࠼ࡽࢀࡓࡇࡽࡇ ࢀ ࢆ 3,1653UHYRWHOOD LQWHUPHGLDQLJUHVFHQFH VSHFLILF UHSHDW ࡋ ࡓ ࠋ 3,165 ࢆ ᚋ 3UH LQWHUPHGLD ࡢ᳨ฟࠊྠᐃ⏝࠸ࡿࡇࡀ࡛ࡁࡿ⪃ ࠼ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 3LQWHUPHGLD ࡣࡢṑ࿘ཎ⳦␗࡞ࡗ࡚ዷፎᛶ ṑ࿘⅖ࠊᛮᮇᛶṑ࿘⅖ࡢ㛵㐃ࡀ▱ࡽࢀ࡚࠸ࡿࡀࠊ ࡑࡢᶵᵓࡣᮍゎ࡛᫂࠶ࡿࠋᮏᨭࢆᇶᚓࡽࢀࡓࢤ ࣀ࣒ሗࢆά⏝ࡍࡿࡇ࡛ࠊᮏ⳦ࡢཎᛶࠊཎࣉ ࣟࢸ࣮ࢮ࣭ࢻ࣊ࢪࣥࡢ⭷㍺㏦࣭ศἪᶵᵓࡢẚ㍑ ࢆ⾜࠺ࡇࡀ࡛ࡁࡓࠋᮏᨭᚋࡢᒎ㛤ࡋ࡚ࠊᚓࡽ ࢀࡓࢤࣀ࣒ሗࢆࡶཎ㑇ఏᏊ➼ࡢ㑇ఏᏊ≉␗ ⓗ࡞ኚ␗ᰴࡢసᡂࢆ┠ᣦࡋ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 1DLWR0HWDO'1$5HVHDUFKLQSUHVV - 454 - බເᨭㄢ㢟㸸2013 ᖺᗘ ࢪே✀ࡢ㣗⭠ෆࣇ࣮ࣟࣛᗣ㛵ࡍࡿㄪᰝ◊✲ ᇶ┙◊✲㸦㹀㸧 ୰ᒣ 㑻࣭ᕞᏛᏛ㝔㎰Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 ࠕ㣗యᗣࠖ㛵ࡍࡿࣛࣇࢧ࢚ࣥࢫࡢ 㛵ᚰࡀ᪥ࠎ㧗ࡲࡗ࡚ࡁ࡚࠸ࡿ᪥ࠊከẸ᪘ࡀΰᅾࡍ ࡿࢪࡣࠊࣘࢽ࣮ࢡ࡛ᗣຠᯝࡢ㧗࠸㣗ᩥࡢᐆ ᗜ࡛ࡶ࠶ࡾࠊ ࠕ㣗యᗣࠖࡢᏛࣇ࣮ࣝࢻࡋ ࡚ὀ┠ࡉࢀ࡚࠸ࡿࠋ୍᪉ࠊ㏆ᖺࡢ⭠ෆ⣽⳦ᏛࡢⓎᒎ ࡼࡾࠊ500㹼1000 ✀ࡢ⣽⳦ࡽᵓᡂࡉࢀࡿ⭠ෆࣇ ࣮ࣟࣛࡣࠊ㣗ᗣࢆ⧅ࡄࣥࢱ࣮ࣇ࢙࣮ࢫࡋ࡚ 㠀ᖖ㔜せ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿࡇࡀศࡗ࡚ࡁ ࡓࠋࡑࡇ࡛ࠊᡃࠎࡣࠊࢪㅖእᅜ࣭ィ 10 ࣨᅜࡢ◊ ✲ ࢢ ࣝ ࣮ ࣉ ඹ ྠ ࡛ Asian Microbiome Project (AMP)ࢆ❧ࡕୖࡆࠊࢪྛᆅᇦ࠾ࡅࡿ㣗⭠ෆ ⣽⳦ྀᗣ≧ែࡘ࠸࡚ࠊヲ⣽ࡘ⥙⨶ⓗㄪᰝ ࡋࠊᇶ┙ࢹ࣮ࢱ࣮࣋ࢫࢆᵓ⠏ࡋ࡚࠸ࡿࠋࡑࡋ࡚ࠊࡑ ࡢࢹ࣮ࢱ࣮࣋ࢫࢆከḟඖⓗゎᯒࡋࠊே✀㣗⭠ ෆ⣽⳦ྀᗣࡢ㛫ࡢ㛵㐃ᛶࢆᩘከࡃぢฟࡍࡇࢆ ┠ᶆࡋ࡚࠸ࡿࠋࡇࢀࡽࡢ▱ぢࡣࠊࢪⓎࡢ⭠ෆ ⣽⳦ࢆࡍࡿᗣ㣗ရࡸ་⸆㛤Ⓨࡶࡁࡃ㈉⊩ࡍ ࡿᮇᚅࡉࢀࡿࡇࢁ࡛࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 AMP ࡛ࡣࠊ➨୍ᮇ◊✲㸦Phase I study㸧ࡋ࡚ࠊ ྛᅜ≉᭷ࡢ㣗ࢆᦤྲྀࡍࡿᶵࡀࡼࡾከ࠸ᮇᚅࡉࢀ ࡿඣ❺㸦㸵ṓ㹼㸯㸯ṓ㸧ࢆᑐ㇟ࠊ㣗⭠ෆ⣽⳦ྀ ࡢㄪᰝࢆ⾜ࡗࡓࠋࡑࡋ࡚ 16S rRNA 㑇ఏᏊࡼࡿ⳦ ྀゎᯒࡣ⛉◊㈝ᇶ┙◊✲ B ᾏእᏛ⾡ㄪᰝࡼࡾ⊂⮬ ⾜࠸ࠊ303 ྡࡢ⳦ྀࣉࣟࣇࣝࢹ࣮ࢱࢆᚓࡓࠋ ᮏࢤࣀ࣒ᨭ◊✲࡛ࡣࠊࡇࡢ Phase I study ࡛ᑐ㇟ ࡋࡓࢧࣥࣉࣝࡢ୰ࡽ≉ᚩⓗ࡞⳦ྀࣉࣟࣇࣝ ࢆ᭷ࡍࡿ 36 ࢧࣥࣉࣝ㸦ࣂࣥࢥࢡ㸵ࢧࣥࣉࣝࠊࢥࣥࢣ ࣥ㸴ࢧࣥࣉࣝࠊࢪࣙࢢࢪࣕ࢝ࣝࢱ㸳ࢧࣥࣉࣝࠊࣂࣜ 㸳ࢧࣥࣉࣝࠊ⚟ᒸ㸷ࢧࣥࣉࣝࠊ㡑ᅜ㸲ࢧࣥࣉࣝ㸧ࢆ 㑅ᢥࡋࠊࢩࣙࢵࢺ࣓࢞ࣥࢱࢤࣀ࣒ゎᯒࢆ⾜ࡗ࡚࠸ࡓ ࡔ࠸ࡓࠋ ྛ⣅౽ࢧࣥࣉࣝࡼࡾᢳฟࡋࡓ⣽⳦ࢤࣀ࣒ DNA ࢆ ⢭〇ࡋࠊIllumina Nextera Kit ࡛ࢩ࣮ࢣࣥࢫ⏝ࡢ DNA ᩿∦ࢆㄪᩚࡋࠊIllumina HiSeq ࡼࡾ 150 bp ࡢ࢚࣌ࣥࢻࢩࣙࢵࢺ࢞ࣥࢩ࣮ࢣࣥࢫࢆ⾜ࡗࡓࠋ⤖ ᯝࠊྛࢧࣥࣉࣝ࠶ࡓࡾᖹᆒ⣙ 10Gb ࡢ㓄ิࢆᚓࡓࠋ ࣄࢺࢤࣀ࣒┦ᙜࡍࡿ㓄ิࡣࠊࣄࢺࢤࣀ࣒㓄ิࣜࣇ ࣞࣥࢫࢹ࣮ࢱ࣮࣋ࢫ hg38 ᑐࡋ࡚ࠊᚓࡽࢀࡓ㓄ิ ࢆ Bowtie ࠾ࡼࡧ BMTagger ࡢࣉࣟࢢ࣒ࣛࢆ⏝࠸࡚ ࣐ࢵࣆࣥࢢࡉࡏࡿࡇ᳨࡛ฟࡋࠊ㝖ཤࡋࡓࠋᚓࡽࢀ ࡓ࣮࣌㓄ิࢆ㐃⤖ࡋࠊᖹᆒ㛗 170 base ࡢ᩿∦ࡢ㓄 ิࡀྛࢧࣥࣉࣝ࠶ࡓࡾ 11.1r8,5 M ᮏࢆᚓࡓࠋࡑࢀࡽ ࢆ Bowtie2 ࡛࣐ࢵࣆࣥࢢࠊPlatanus-UD ࡛ࢵࢭࣥ ࣈࣝࡋ࡚ࠊྛࢧࣥࣉࣝ࠶ࡓࡾ⣙ 40 ᮏ㸦ᖹᆒ㛗 671 bp㸧ࡢ contig ࢆᚓࡓࠋMetaGeneMark ࡼࡿண 㑇ఏᏊᩘࡣྛࢧࣥࣉࣝ࠶ࡓࡾ⣙ 55 ࡛࠶ࡗࡓࠋḟࠊ ࡑࢀࡽࡢ㓄ิࢆ⏝࠸࡚ࠊKEGG Gene Blast ࢆ⾜ࡗࡓࠋ ࡑࡢ⤖ᯝࠊ⣙ 70㸣ࡀࣄࢵࢺࡋࡓࠋ᭱⤊ⓗࠊKEGG ゎᯒࡽ㑇ఏᏊࡢᶵ⬟࢝ࢸࢦ࣮ࣜࡈࡢᏑᅾẚࢹ࣮ ࢱࢆᚓࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᡃࠎࡀᇶ┙◊✲㸦B㸧ࡼࡾ⊂⮬ゎᯒࡋࡓ AMP Phase I ࠾࠸࡚ࢧࣥࣉࣜࣥࢢࡋࡓࢪ㸳ࢣᅜࡢ ᑠᏛඣ❺ 303 ྡࡢ 16S rRNA ࡢ㓄ิゎᯒ⤖ᯝࡽࠊ ࢪඣ❺ࡢ⭠ෆ⣽⳦ྀࡀࡁࡃ㸰ࡘࡢࢱࣉศࡅ ࡉࢀࡿࡇࢆぢฟࡋ࡚࠸ࡿࠋ୍ࡘࡣࣉࣞ࣎ࢸࣛᒓ⣽ ⳦ࢆඃ༨⳦ࡍࡿ P ࢱࣉࠊࡶ࠺୍ࡘࡣࠊࣂࢡࢸࣟ ࢹࢫᒓ࠾ࡼࡧࣅࣇࢬࢫ⳦ࢆඃ༨⳦ࡍࡿ BB ࢱ ࣉ࡛࠶ࡿࠋࡇࢀࡽࡣࠊḢᕞࡢ MtetaHIT ࡢࢢ࣮ࣝ ࣉࡀⓎぢࡋࡓࣄࢺ⭠ෆ⣽⳦ࡢࡁ࡞㸱ࡘࡢࢱࣉ 㸦࢚ࣥࢸࣟࢱࣉ㸧┦ᙜࡍࡿ⪃࠼ࡽࢀࡿࠋ ᅇࡢࢤࣀ࣒ᨭ◊✲࡛ᚓࡽࢀࡓ࣓ࢱࢤࣀ࣒ゎᯒ ࡽࠊP ࢱࣉ࠾ࡼࡧ BB ࢱࣉࡢ⣽⳦ྀࢥ࣑ࣗࢽ ࢸ࣮ࡀࢥ࣮ࢻࡍࡿࢱࣥࣃࢡ㉁ࡢᶵ⬟ᵓᡂࡢ≉ᚩࢆ ぢฟࡍࡇࡀ࡛ࡁࡓࠋ≉⯆῝࠸≉ᚩࡋ࡚ࠊBB ࢱࣉࡣࠊ⫹Ồ㓟⏕ྜᡂ㓝⣲㑇ఏᏊࡢᏑᅾẚࡀ㧗 ࡗࡓࠋࡲࡓࠊP ࢱࣉࡣࠊ࣑࣮ࣛࢮࡸ࣌ࢡࢳ ࢼ࣮ࢮ㛵㐃ࡢ㑇ఏᏊࡢᏑᅾẚࡀ㧗ࡗࡓࠋBB ࢱࣉ ࡣ᪥ᮏࠊ୰ᅜࠊྎ‴ከࡃࠊࣥࢹ࢝⡿⏤᮶ࡢ㞴 ᾘᛶࢹࣥࣉࣥࢆከࡃᦤࡗ࡚࠸ࡿࣥࢻࢿࢩࡸࢱ ࡢᏊࡶ㐩ẚ࡚⭠⟶࠾ࡅࡿ⬡㉁ࡢ⃰ᗘࡀ㧗 ࡃࠊ⫹Ồ㓟ࡢ⃰ᗘࡶ㧗ࡃ࡞ࡾࠊࣉࣞ࣎ࢸࣛ࡞⫹Ồ 㓟ᑐࡍࡿ⪏ᛶࡀప࠸⣽⳦ࢆಖᣢ࡛ࡁ࡞ࡃ࡞ࡗ࡚࠸ ࡿࡢ࡛ࡣ࡞࠸᥎ᐹࡉࢀࡿࠋ ࡲࡓࠊP ࢱࣉ BB ࢱࣉࢥ࣮ࢻࡉࢀࡿࢱࣥ ࣃࢡ㉁ࡢᶵ⬟ᵓᡂࢆయẚ㍑ࡍࡿࠊ୧⪅ࡢ㛵ಀࡀࠊ ࣓ࣜ࢝ேࣇࣜ࢝ேࡢ㛵ಀ(Yatsunenko et al., Nature 2012)㢮ఝࡋ࡚࠸ࡓࠋࡘࡲࡾᮾ༡ࢪ ࠾࠸࡚ࡶḢ⡿ࡢᙳ㡪ࡀ⭠ෆࣇ࣮ࣟࣛ⌧ࢀ࡚࠸ࡿ ⪃࠼ࡽࢀ㠀ᖖ⯆῝࠸ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 16S rRNA ࡢ㓄ิሗࡽࡣᚓࡿࡇࡀ࡛ࡁ࡞ ࡗࡓ⭠ෆ⣽⳦ྀࡢᶵ⬟ሗࢆヲ⣽ᚓࡿࡇࡀ࡛ࡁ ࡓࠋ≉ࠊᡃࠎࡀ㐍ᒎࡉࡏ࡚࠸ࡿࢪேࡢ⭠ෆࣇ ࣮ࣟࣛࡢẚ㍑◊✲࡛ࡣࠊ㣗ᗣࡢࣥࢱ࣮ࣇ࢙࣮ ࢫࡋ࡚ࡢ⭠ෆࣇ࣮ࣟࣛࡢᶵ⬟ࢆ⌮ゎࡍࡿࡇࢆ᭱ ⤊┠ᶆࡋ࡚࠸ࡿࡇࡽࠊᅇࡢ࣓ࢱࢤࣀ࣒◊✲ ࡼࡾࡁࡃࢦ࣮ࣝ㏆࡙ࡃࡇࡀ࡛ࡁࡓ⪃࠼ࡿࠋ ᚋࡼࡾヲ⣽ࢹ࣮ࢱࢆゎᯒࡋࠊ࡛ࡁࢀࡤ᪂ࡓࢧ ࣥࣉࣝࡑࡢ࣓ࢱࢤࣀ࣒ࢹ࣮ࢱࡶ㞟ࡋ࡚࠸ࡃࡇ ࡛ࠊࢪࡢከᵝ࡞㣗ࡀࠊࡑࢀࡒࢀᐟࡢࡼ࠺ ࡞ᙳ㡪ࢆ࠼ࠊࡑࡢ⤖ᯝࡢࡼ࠺ࢪேࡢᗣ ᙳ㡪ࢆཬࡰࡋ࡚࠸ࡿࠊᗈࡃࠊ῝ࡃࠊṇ☜⌮ゎ ࡋ࡚࠸ࡁࡓ࠸⪃࠼ࡿࠋ 㸺ᨭᡂᯝබ⾲ㄽᩥࣜࢫࢺ㸼 ᮾె㑣Ꮚ, ୰ᒣ㑻, ⭠ෆ⣽⳦Ꮫ㞧ㄅ 29, 135-144 (2015). - 455 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࢪே✀ࡢ㣗⭠ෆࣇ࣮ࣟࣛᗣ㛵ࡍࡿㄪᰝ◊✲ ᇶ┙◊✲㸦㹀㸧 ୰ᒣ 㑻࣭ᕞᏛᏛ㝔㎰Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 21 ୡ⣖ࡢᖥ㛤ࡅࡶ⇿Ⓨⓗ㐍ᒎࡋࡓࢤࣀ࣒ ⛉Ꮫࡢᢏ⾡㠉᪂ࡣࠊ⭠ෆඹ⏕⳦Ꮫࡶከ࡞ࡿἼཬ ຠᯝࢆཬࡰࡋࠊ⭠ෆඹ⏕⳦ᐟࡢᗣࡢ㛵ಀࡘ ࠸࡚ḟࠎ࣓ࢫࢆධࢀ࡚࠸ࡿࠋ≉᭱㏆࡛ࡣࠊ1000 ✀ࡢࡰࡿゝࢃࢀࡿඹ⏕⳦⩌࠾࠸࡚ࠊࡢ⳦ࡀ ࡢࡼ࠺ᐟᙳ㡪ࢆ࠼࡚࠸ࡿࠊヲ⣽࡞ࢹ࣮ ࢱࡀ♧ࡉࢀࡿࡼ࠺࡞ࡗ࡚ࡁࡓࠋ ࡇ ࡢ ࡼ ࠺ ࡞ ௦ ⫼ ᬒ ࡢ ୰ ࠊ ⡿ ᅜ NIH ᑟ ࡢ Human Microbiome Project(HMP) ࠊ Ḣ ᕞ ࡛ ࡣ Metagenomics of Human Intestinal Tract (MetaHIT)ࠊከேᩘࡢ⭠ෆࣇ࣮ࣟࣛࢹ࣮ࢱࢆ㞟 ࡍࡿつᶍࣉࣟࢪ࢙ࢡࢺࡀ㐍⾜ࡋ࡚࠸ࡿࠋࡑࡋ࡚ࠊ ࢪ࡛ࡶࠊ⮬㌟ࡢ⭠ෆࣇ࣮ࣟࣛࡢࢹ࣮ࢱࢆ⮬ศࡓ ࡕࡢᡭࡼࡾ㞟ࡋࡼ࠺࠸࠺ᶵ㐠ࡀ㧗ࡲࡾࠊ2009 ᖺࢪங㓟⳦Ꮫ㐃ྜࡢ࣓ࣥࣂ࣮ࡀ୰ᚰ࡞ࡾࠊ Asian Microbiome Project (AMP) ࢆ ጞ ື ࡋ ࡓ ࠋ Phase I study ࡛ࡣࠊࡑࢀࡒࢀࡢᅜࡢ㣗ᩥࢆᫎࡍ ࡿᐙᗞ㣗ࢆࡍࡿᑠᏛඣ❺ࢆᑐ㇟ࠊ5 ࣨᅜ 10 㒔 ᕷ࡛ㄪᰝࢆ⾜ࡗࡓ㸦Nakayama et al., 2015. Sci. Rep㸧ࠋィ 303 ྡࡢ⣅౽ࢧࣥࣉࣝࡘ࠸࡚࣓ࢱ 16S rRNA ゎᯒࢆ⾜࠸ࠊᚓࡽࢀࡓ⣽⳦⤌ᡂࢹ࣮ࢱࢆᇶࠊ ᡂศศᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊ⯆῝࠸ࡇࠊ ࢪேࡢ⭠ෆࣇ࣮ࣟࣛࡣࡁࡃࡘࡢࢱࣉࢡ ࣛࢫࢱࣜࣥࢢࡉࢀࡿࡇࡀุ᫂ࡋࡓࠋ୍ࡘࡣࠊࣉࣞ ࣎ࢸࣛᒓ⣽⳦ࢆࢥࡍࡿ P ࢱࣉ࡛ࠊࣥࢻࢿࢩ ࢱከࡃぢࡽࢀࡿࠋ୍᪉ࠊ᪥ᮏࠊ୰ᅜࠊྎ‴ ࡢඣ❺ࡣࣅࣇࢬࢫ⳦ࣂࢡࢸࣟࢹࢫᒓࢆ㇏ᐩ ᭷ࡍࡿ⩌ᒓࡋ BB ࢱࣉྡࡋࡓࠋ ⥆࠸࡚ 2013 ᖺᗘࠊࢤࣀ࣒ᨭ◊✲ࡢᨭࢆཷࡅࠊ ୖグ 303 ࢧࣥࣉࣝࡽࠊ≉ᚩⓗ࡞⣽⳦⤌ᡂࢆ♧ࡋࡓ 36 ࢧࣥࣉࣝࢆ㑅ᢥࡋࠊࢩࣙࢵࢺ࣓࢞ࣥࢱࢤࣀ࣒ゎᯒ ࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊBB ࢱࣉࡣ⫹Ồ㓟⬺ᢪྜ㓝 ⣲㑇ఏᏊࡀࡼࡾ㇏ᐩ࡛ࠊP ࢱࣉࡣȘ࣑࣮ࣛࢮ ࡢ㑇ఏᏊࡀࡼࡾ㇏ᐩ࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋྛᅜࡢ 㣗⩦័ࣥࢣ࣮ࢺㄪᰝ⤖ᯝࢆ↷ྜࡋ࡚ࠊP ࢱࣉࡢ ேࡣࣥࢹ࢝⡿ࡸࢪࣕ࣡ࢽ࢝⡿࡞ࢪ࣏ࣕࢽ࢝⡿ ẚࡿ㞴ᾘᛶࢹࣥࣉࣥࢆከࡃྵࡴ⡿ࡢᦤྲྀ㔞 ࡀከࡃࠊࡑࡢࡓࡵࣉࣞ࣎ࢸࣛᒓ࡞Ș࣑࣮ࣛࢮࢆ ᭷ࡍࡿ⣽⳦ࡀከࡃ࡞ࡗ࡚࠸ࡿࡢᑐࡋࠊBB ࢱࣉ࡛ ࡣࠊ㞴ᾘᛶࢹࣥࣉࣥࡸ㣗≀⧄⥔ᦤྲྀ㔞ࡀᑡ࡞ࡃࠊ ⭠⟶ࡢ⫹Ồ㓟㔞ࡀ㧗ࡃ࡞ࡾ⫹Ồ㓟ឤཷᛶࡢࣉࣞ࣎ࢸ ࣛᒓ⣽⳦ࡀ⏕ᜥ࡛ࡁ࡞ࡃ࡞ࡗ࡚࠸ࡿࠊ࠸࠺సᴗ௬ ㄝࢆ❧࡚ࡿ⮳ࡗ࡚࠸ࡿࠋ 2014 ᖺᗘࡣࠊP ࢱࣉࡢከ࠸ࣥࢻࢿࢩ BB ࢱࣉࡢከ࠸᪥ᮏ࡚ฟ⏕ࡋࡓஙᗂඣࢆᑐ㇟ ⣅౽⣽⳦ྀࡢゎᯒࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊኚ⯆῝ ࠸ࡇࠊP ࢱࣉࡢࣥࢻࢿࢩࡢฟ⏕ඣࡣ㞳ங ᚋ୍᪦ࣂࢡࢸࣟࢹࢫࡀ㇏ᐩ࡞ࣇ࣮ࣟࣛࢆᵓ⠏ࡍࡿ ࡀࡑࡢᚋࡲࡶ࡞ࡃ P ࢱࣉ㌿ࡍࡿࡇࡀุ᫂ࡋ ࡓࠋࡑࡋ࡚ࠊࡇࡢᐇࡽᡃࠎࡣࠊஙᗂඣࡢ⣅౽⣽ ⳦ྀࡢ⤒ኚࢆ㣗ㄪᰝࠊࢤࣀ࣒ᨭ◊✲ࡼ ࡿ࣓ࢱࢤࣀ࣒ゎᯒࢆࣜࣥࢡࡉࡏ࡚㐍ᒎࡉࡏࡿࡇ࡛ࠊ P BB ࡢ⭠ෆࣇ࣮ࣟࣛࢱࣉࡀᵓ⠏ࡉࢀ࡚࠸ࡃࢲ ࢼ࣑ࢬ࣒ࡢ⌮ゎ㏕ࡾࡓ࠸⪃࠼࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࡇࢀࡲ࡛ࡢ AMP ࠾ࡅࡿ◊✲ᡂᯝࡽࠊࢪ ேࡢ⭠ෆࣇ࣮ࣟࣛࡣࠊ᪥ᮏࡸ୰ᅜከ࠸ࣅࣇࢬࢫ ⳦࣭ࣂࢡࢸࣟࢹࢫᒓ⣽⳦ࢆ㇏ᐩྵࡴ BB ࢱࣉ ࠊࣥࢻࢿࢩࡸࢱከ࠸ࣉࣞ࣎ࢸࣛᒓ⣽⳦ࢆ ㇏ᐩྵࡴ P ࢱࣉࡢ㸰ࡘࡢࢱࣉࡁࡃศࢀ ࡿࡇࡀ♧ࡉࢀ࡚࠸ࡿࠋࡑࡋ࡚ࠊP ࢱࣉ BB ࢱ ࣉࡣ㞳ஙᚋࡲࡶ࡞ࡃࡢᗂඣᮇᙧᡂࡉࢀࡿࡇࡶ ุ᫂ࡋࡓࠋᮏᨭ◊✲࡛ࡣࠊ᪥ᮏࣥࢻࢿࢩࡢ ẕぶஙᗂඣࡢ᭶࣭ᖺ㱋ࡈࡢ⣅౽ࢧࣥࣉࣝࡢࢩࣙ ࢵࢺ࣓࢞ࣥࢱࢤࣀ࣒ゎᯒࢆ⾜࠸ࠊձẕぶᏊࡢ㛫ࡢ ⭠ෆ⣽⳦ྀࡢ┦㛵ࠊղࣅࣇࢬࢫࣇ࣮ࣟࣛࡽࣂࢡ ࢸࣟࢹࢫࣇ࣮ࣟࣛࢆ⤒⏤ࡋ࡚ࣉࣞ࣎ࢸࣛࣇ࣮ࣟࣛ ࡀᙧᡂࡉࢀࡿࡲ࡛ࡢࠊ⣽⳦ࢥ࣑ࣗࢽࢸ࣮ࡢᶵ⬟ኚ ࠊճ᪥ᮏࣥࢻࢿࢩࡢஙᗂඣࡢ⏕ᚋࡢ⭠ෆࣇ ࣮ࣟࣛኚ㑄ࡢ┦㐪ࡑࢀࢆ⏕ࡴせᅉࠊ௨ୖࢆ᫂ࡽ ࡍࡿࡇࢆィ⏬ࡋ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ୖグࢩࣙࢵࢺ࣓࢞ࣥࢱࢤࣀ࣒ゎᯒࢆ⌧ᅾᐇ୰࡛ ࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࠕ㣗యᗣࠖ㛵ࡍࡿࣛࣇࢧ࢚ࣥࢫࡢ㛵 ᚰࡀ᪥ࠎ㧗ࡲࡗ࡚ࡁ࡚࠸ࡿ᪥ࠊከẸ᪘ࡀΰᅾࡍࡿ ࢪࡣࠊࣘࢽ࣮ࢡ࡛ᗣຠᯝࡢ㧗࠸㣗ᩥࡢᐆᗜ ࡛ࡶ࠶ࡾࠊ ࠕ㣗యᗣࠖࡢᏛࣇ࣮ࣝࢻࡋ࡚ ὀ┠ࡉࢀ࡚࠸ࡿࠋ୍᪉ࠊ㏆ᖺࡢ⭠ෆ⣽⳦ᏛࡢⓎᒎ ࡼࡾࠊ500㹼1000 ✀ࠊ100 ಶࡢ⣽⳦ࡽᵓᡂࡉࢀ ࡿゝࢃࢀࡿ⭠ෆࣇ࣮ࣟࣛࡣࠊ㣗ᗣࢆ⧅ࡄࣥ ࢱ࣮ࣇ࢙࣮ࢫࡋ࡚㠀ᖖ㔜せ࡞ᙺࢆᯝࡓࡋ࡚࠸ ࡿࡇࡀศࡗ࡚ࡁࡓࠋᮏ◊✲࡚ࠊ↓⳦≧ែࡽ ࡇࡢࡼ࠺࡞㧗ᗘ࡞⏕ែ⣔ࡀࣄࢺࡢ⭠⟶࡚ᙧᡂࡉࢀ ࡚࠸ࡃ㐣⛬ࢆࢤࣀ࣒࡛ࣞ࣋ࣝࣔࢽࢱࣜࣥࢢࡍࡿࡇ ࡼࡾࠊࡑࡢ⏕ែ⣔ࢆᅄḟඖ⌮ゎࡍࡿࡇࡀྍ⬟ ࡞ࡾࠊࡉࡽࡣᐟࡢᡂ㛗ࡶ࠼ࡿᙳ㡪ࡶከࡃࡢ ሗࡀᚓࡿࡇࡀ࡛ࡁࡿᮇᚅࡉࢀࡿࠋ࠶㸰ࣨ᭶ 㛫࡛ࡢࢹ࣮ࢱゎᯒࡢ㐍ᒎᙉࡃᮇᚅࡍࡿࠋ 㸺ᨭᡂᯝබ⾲ㄽᩥࣜࢫࢺ㸼 ↓ࡋ - 456 - බເᨭㄢ㢟㸸2012ࠥ2013 ᖺᗘ ࢤࣀ࣒ゎᯒࡼࡿᐙ᪘ᛶ IgA ⭈ࡢཎᅉ㑇ఏᏊࡢ᥈⣴ ᇶ┙◊✲㸦㹁㸧 ᡂ⏣ ୍⾨࣭᪂₲ᏛᏛ㝔་ṑᏛ⥲ྜ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⭈⏕᳨࡛☜ᐃデ᩿ࡋࡓ ,J$ ⭈ᝈ⪅࠾࠸࡚ࠊ୍ぢ ᏙⓎᛮࢃࢀࡿ࡛ࡶ⣙ 㸣ᒀ␗ᖖࡸ⭈ ࡢᐙ᪘Ṕࡀほᐹࡉࢀࠊ,J$ ⭈ࡼࡿ⭈࡛⫗ぶ ࢆࢻࢼ࣮ࡍࡿ⭈⛣᳜ࢆ⾜࠺ࠊ⛣᳜┤๓ࡢࢻࢼ࣮ ⭈⣒⌫య ࠥ ࡢ㢖ᗘ࡛ ,J$ ࡢ⣒⌫యỿ╔ࡀ ほᐹࡉࢀࡿࠋ,J$ ⭈ࡢⓎ㑇ఏせᅉࡀ㛵ࡋ࡚࠸ ࡿ⪃࠼ࡽࢀࡿࠋ ᮏ◊✲ࡢ┠ⓗࡣࠊᐙ᪘ᛶ ,J$ ⭈ࡢᝈឤཷᛶ㑇 ఏᏊࢆྠᐃࡋࠊࡑࢀࢆ㏻ࡋ࡚ᮏࡢⓎᶵᗎࢆࡼࡾ ヲ⣽⌮ゎࡍࡿࡇ࡛࠶ࡿࠋᐙ᪘ᛶ ,J$ ⭈ࡢཎᅉ ࢆ᫂ࡽࡍࡿࡓࡵࠊከᩘࡢᐙ⣔ࢆᑐ㇟ࡋࡓ㐃 㙐ゎᯒࡀ⾜ࢃࢀ࡚ࡁࡓࡀࠊ⌧ᅾࡲ࡛㈐௵㑇ఏᏊࡣ ྠᐃࡉࢀ࡚࠸࡞࠸ࠋ㏆ᖺࠊ㧗㏿ࢩ࣮ࢣࣥࢧࡼࡿ 㔞ࡢ㑇ఏᏊ㓄ิሗࡽᐙ᪘ᛶᕼᑡᝈࢆ୰ᚰࡋ ࡓᝈ㑇ఏᏊࡢゎ᫂ࡀ㐍ࢇ࡛࠸ࡿࠋᅇࠊᐙ᪘ᛶ ,J$ ⭈ࡶࡇࡢゎᯒ᪉ἲࢆヨࡳࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᖺᗘࡣࠊᐙ᪘ᛶ ,J$ ⭈ ᐙ⣔ࡘ࠸࡚ ྡࠊ ᖺᗘࡣ ᐙ⣔ࡘ࠸࡚ࠊ ྡࡢ࢚ࢡࢯ࣮࣒ゎ ᯒࡢᨭࢆ࠸ࡓࡔ࠸ࡓࠋ ࢀ࡚࠾ࡾࠊ,J$ ⭈ࡢⓎ㛵ࡍࡿྍ⬟ᛶࡀ࠶ࡿࠋ ᖺᗘࡣࠊẚ㍑ⓗᑠつᶍ࡞ ᐙ⣔ࡘ࠸࡚ࠊ࢚ ࢡࢯ࣮࣒ゎᯒ㐃㙐ゎᯒࢆ⤫ྜࡋࠊೃ⿵㑇ఏᏊኚ␗ ࡢྠᐃࢆヨࡳࡓࠋ࢚ࢡࢯ࣮࣒ゎᯒࡼࡗ࡚ᚓࡽࢀࡓ 619LQVGHO ࡽࠊ࣑ࣀ㓟⨨ࢆక࠸ࠊ㢖ᗘࡀపࡃ 㸦+XPDQ*HQHWLF9DULDWLRQ'DWDEDVH ࡛ ௨ୗ㸧 ࠊ ࡑࡋ࡚⨯ᝈ⪅ࡢࡳㄆࡵࡽࢀࡿ 619LQVGHO ࢆྛᐙ ⣔ẖ⤠ࡾ㎸ࢇࡔࠋࡲࡓࠊ࢚ࢡࢯ࣮࣒ゎᯒ࡛ᚓࡽࢀ ࡓ ᐙ⣔ࡢ 619 ࢹ࣮ࢱࢆ⏝࠸࡚ࠊࢤࣀ࣒㐃㙐ゎᯒ 㸦OLQNGDWDJHQPHUOLQ㸧ࢆ⾜ࡗࡓࠋࢤࣀ࣒㐃㙐ゎ ᯒ࡛ࡣࠊࣃ࣓ࣛࢺࣜࢵࢡゎᯒ࡛ /2' ࢫࢥ㸼 ࢆ♧ ࡍ 㡿ᇦ㸦SSTTT㸧 ࡀ᳨ฟࡉࢀࡓࠋྛᐙ⣔㛫࡛ඹ㏻ࡋ࡚ㄆࡵࡽࢀࡿ 619LQVGHO ࢆࣜࢫࢺࡋ㸦YDULDQWV㸧ࠊࡉࡽ㐃 㙐ゎᯒࡢࣆ࣮ࢡ㡿ᇦᏑᅾࡍࡿ YDULDQW ⤠ࡾ㎸ࢇ ࡔ㸦TT㸧 ࠋ ᖺᗘ㸸ᐙ᪘ᛶ ,J$ ⭈ ᐙ⣔ࡢ㐃㙐ゎᯒ 7B ᐙ⣔ 1B ᐙ⣔ ᖺᗘ㸸࢚ࢡࢯ࣮࣒ゎᯒࡀ⾜ࢃࢀࡓᐙ᪘ᛶ ,J$ ⭈ ᐙ⣔ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᖺᗘࡣࠊ,J$ ⭈ࡀ ࠥ ྡ௨ୖㄆࡵࡽࢀࡿ ᐙ ⣔ࢆᑐ㇟ࡋ࡚࢚ࢡࢯ࣮࣒ゎᯒࡀ⾜ࢃࢀࡓࠋࡇࢀ ඛ❧ࡕᨭ౫㢗⪅ࡽࡣ ,J$ ⭈ ྡࢆྵࡴᐙ⣔ 㸦1B㸧ࡢ࢚ࢡࢯ࣮࣒ゎᯒࢆ⾜࠸ࠊ࢚ࢡࢯࣥࡢ㓄 ิሗࡽࣇࣝࢱࣜࣥࢢࢆ⾜࠸ࠊ⨯ᝈ⪅ࡢࡳඹ ㏻ࡍࡿ ࡢ YDULDQW ࡀ㑅ูࡉࢀࡓࠋୖᅗࡢ 1B ᐙ⣔࡛ࡣ ࠊ7B ᐙ⣔࡛ࡣ ࡢ YDULDQW ࡀ㑅ูࡉ ࢀࡓࡀࠊ ᐙ⣔࡛ඹ㏻ࡍࡿኚ␗ࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ 1B ᐙ⣔࡛ࡣࠊ(($S)< ࡣ㐃㙐ゎᯒ࡛ /2' ࢫ ࢥࡀ᭱ࡶ㧗࠸㡿ᇦෆㄆࡵࡽࢀࠊᐙ⣔ෆ FRVHJUHJDWLRQ ࡀ୍⮴ࡋࡓࠋࡉࡽࡢ )< ᐙ⣔࠾࠸࡚ (($ ኚ␗ࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜࠸ࠊ ࡣ␗࡞ࡿኚ␗㸦5:1.(*㸧ࢆㄆࡵࠊ ᪥ᮏேࢹ࣮ࢱ࣮࣋ࢫẚ㍑ࡋࠊ(($ ኚ␗ࡣᐙ᪘ᛶ ,J$ ⭈᭷ពከࡃㄆࡵࡽࢀࡓࠋ(($ ࡣึᮇ࢚ࣥࢻࢧ ࢺ࣮ࢩࢫᑠ⬊㛫ࡢ⼥ྜࡽࢯ࣮ࢸࣥࢢᚲ㡲ࡢ ⺮ⓑ࡛࠶ࡿࠋ,J$ ศᏊࡢࢺࣛࣥࢫࢧࢺ࣮ࢩࢫࡶ㛵 ࡋ࡚࠾ࡾࠊ⢓⭷ච␗ᖖࡽ ,J$ ⭈ࡢⓎ㛵 ࡍࡿྍ⬟ᛶࡀ࠶ࡿࠋࡲࡓࠊ7B ᐙ⣔࡛ࡢೃ⿵㑇ఏ Ꮚኚ␗⩌ࡣࣜࣥࣃ⌫ඃⓎ⌧ࡍࡿ㑇ఏᏊࡀྵࡲ T ࡣࠊ% ࣜࣥࣃ⌫Ⓨ⌧ࡋࠊᙧ㉁⣽⬊ࡢศ ࢆㄪ⠇ࡍࡿ㑇ఏᏊࡢ 619 ࡀྵࡲࢀ࡚࠾ࡾࠊ࣑ࣀ㓟 ⨨ࡢᙳ㡪ࡶࡁ࠸⪃࠼ࡽࢀࡓࠋ% ࣜࣥࣃ⌫ࡽᙧ ㉁⣽⬊ࡢศ␗ᖖࡀ⢓⭷ච࠼ࡿᙳ㡪ࡘ࠸ ࡚ࠊ,J$ ⭈ࡢⓎࡢ㛵㐃ࢆᝈ⪅ࣜࣥࣃ⌫࡛⌧ᅾࠊ ᳨ウࡋ࡚࠸ࡿࠋ ࢤࣀ࣒ᨭࢆ㏻ࡌ࡚ᚓࡽࢀࡓࡇࢀࡽࡢೃ⿵㑇ఏᏊ ࡘ࠸࡚ᚋࠊᏙⓎࡢኚ␗ࢫࢡ࣮ࣜࢽࣥࢢࡸᶵ⬟ ゎᯒࢆ⾜࠸ࠊ,J$ ⭈ࡢཎᅉ㑇ఏᏊࡋ࡚ࡢド᫂ࢆ㐍 ࡵ࡚⾜ࡃࠋࡇࢀࡼࡾࠊ,J$ ⭈ࡢែࡢゎ᫂ࡀࡉࡽ 㐍ࡴᛮࢃࢀࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡾࠊᐙ᪘ᛶ ,J$ ⭈ࡢ㑇ఏ⫼ᬒࡢᴫ␎ࡀ ᫂ࡽࡉࢀࡘࡘ࠶ࡿࠋ,J$ ⭈ࡢⓎࡣከࡃࡢࢫ ࢸࢵࣉࡀ㛵ࡋ࡚࠾ࡾࠊከᅉᏊᝈࡋ࡚ྛᐙ⣔ࡢ 㑇ఏ⫼ᬒࡣ␗㉁ᛶࡀᙉ࠸ࠋᚲࡎࡋࡶ࡚ࡢ㑇ఏせᅉ ࢆྠᐃ࡛ࡁࡿࡣ㝈ࡽ࡞࠸ࡀࠊᾐ㏱⋡ࡢ㧗࠸ᐙ⣔ࢆ ᑐ㇟ゎᯒࢆ㐍ࡵࡿࡇ࡛ཎᅉ㑇ఏᏊࢆ☜ᐇྠᐃ ࡍࡿࡇࡣྍ⬟⪃࠼ࡽࢀࡿࠋᅜ㝿ඹྠ◊✲࡛ ,J$ ⭈ࡢ *:$6 ࡢゎᯒ⤖ᯝࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀࠊྜࢃࡏ ࡚ᮏ◊✲ࡼࡾ UDUHYDULDQWVPXWDWLRQV ࢆྠᐃࡋࠊ ,J$ ⭈ࡢែࡢㇺゎ᫂ࠊࡲࡓ⸆ྥࡅࡓ⒪ࢱ ࣮ࢤࢵࢺࡢ᥈⣴ࡶᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾࠊⱥᩥㄽᩥࢆ‽ഛ୰ - 457 - බເᨭㄢ㢟㸸2014 ᖺᗘ ࣍ࣟࢤࣀ࣒ゎᯒࡼࡿ IgA ⭈ࡢែゎᯒ⒪ࢱ࣮ࢤࢵࢺ᥈⣴ ᇶ┙◊✲㸦㹀㸧 ᡂ⏣ ୍⾨࣭᪂₲ᏛᏛ㝔་ṑᏛ⥲ྜ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⣒⌫య࣓ࢧࣥࢠ࣒࢘㡿ᇦࡢචࢢࣟࣈࣜࣥ $㸦,J$㸧 ỿ╔ࢆ≉ᚩࡍࡿ ,J$ ⭈ࡣࠊ᭱ࡶ㢖ᗘࡢ㧗࠸ཎⓎ ᛶ⣒⌫య⭈⅖࡛࠶ࡾࠊ៏ᛶ⭈ࡢ࡞ཎᝈࡢ୍ ࡘ࡛ࡶ࠶ࡿࠋࡑࡢⓎࡣࠊ⣽⳦ᢠཎ➼ࡢ⎔ቃᅉᏊ 㑇ఏⓗ⫼ᬒ୧⪅ࡢ㛵ࡀᐃࡉࢀࠊ⌧ᅾࡲ࡛ᮏ ࡢᝈឤཷᛶ㑇ఏᏊࡣ㒊ศⓗ᫂ࡽ࡞ࡗ࡚ࡁࡓ ࡀヲ⣽ࡣ࡛᫂࠶ࡿࠋࡲ࡛⎔ቃᅉᏊ㑇ఏⓗ⫼ᬒ ࡣࠊࡑࢀࡒࢀูಶ◊✲ࡉࢀ࡚ࡁࡓࡓࡵࠊࡑࢀࡽ ࡢ㛵㐃ࢆホ౯ࡍࡿࡇࡣྍ⬟࡛࠶ࡗࡓࠋࡋࡶࠊ ࣄࢺࡣ⮬㌟ࡢ⣽⬊ᩘࡢ ಸ௨ୖࡢᚤ⏕≀⣽⬊ࢆ᭷ ࡋ࡚࠸ࡿࡀࠊࡑࡢ༙ࡣᇵ㣴ྍ⬟࡛࠶ࡾࠊゎᯒ࡛ ࡁ࡞ࡗࡓࠋ ͆࣍ࣟࢤࣀ࣒͇ࡣࠊ࣍ࢫࢺࢤࣀ࣒ࡔࡅ࡛࡞ࡃࠊ ⏕యෆᏑᅾࡍࡿ࡚ࡢ⎔ቃᚤ⏕≀ࢤࣀ࣒ࠊࡉࡽ ࡣ࢚ࣆࢤࣀ࣒ゎᯒࡶྠ⾜࠸ࠊࡑࢀࡽࢆໟᣓⓗ ホ౯ࡍࡿ᪉ἲ࡛ࠊ㏆ᖺࡢゎᯒᢏ⾡ࡢ㐍Ṍࡼࡾึࡵ ࡚ྍ⬟࡞ࡗࡓࡶࡢ࡛࠶ࡿࠋᮏ◊✲ࡣࡇࢀࢆࠊ ,J$ ⭈ࡢⓎ࣭㐍ᒎᶵᗎゎ᫂ᛂ⏝ࡍࡿࠋ ,J$ ⭈࡛ࡣ༙ᩘ௨ୖࡢ࡛⾑Ύ ,J$ ್ࡀୖ᪼ ࡋࠊ,J$ ࡢ⏘⏕࣭௦ㅰ␗ᖖࡀ࠶ࡿࠋࣄࣥࢪ㒊2⤖ ྜᆺ⢾㙐ಟ㣭ࡢ␗ᖖࢆᣢࡘ⢾㙐 ,J$ ศᏊࡣࠊ⮬ ᕫจ㞟ࠊ௦ㅰ㐜ᘏࠊ࣓ࢧࣥࢠ࣒࢘⣽⬊࣭ᇶ㉁ࡢ㧗 ぶᛶ࡞ࡢᛶ㉁ࡀ࠶ࡾࠊᮏࡢཎᅉ┤⤖ࡋ࡚࠸ ࡿ⪃࠼ࡽࢀࡿࠋࡇࡢ,J$ ࡢ2⤖ྜᆺ⢾㙐ࡢᙧᡂ ࡣࠊཎᅉࡣศࡗ࡚࠸࡞࠸ࡀᡥ᱈⢓⭷࡛ ⏕ࡌࠊࡑࢀࡀ ,J$ ࡢ⣒⌫యࡢỿ╔⅖ࢆច㉳ ࡍࡿ㔜せ࡞ࢫࢸࢵࣉ࡛࠶ࡿࡇࠊࡑࡋ࡚⣒⌫య࣓ࢧ ࣥࢠ࣒࢘⣽⬊ഃࡢ␗ᖖࡶ㔜せ࡛࠶ࡿࡇࡀศ ࡗ ࡚ ࠸ࡿࠋ ཱྀᡥ᱈ࡢ␗ᖖࡀᮏⓎ࠾࠸࡚㔜せ࡞ᙺࢆ ᯝࡓࡍࡇࡣࠊ⮫ᗋⓗᐇࡽࡶ᫂ࡽ࡛࠶ࡿࠋᡥ ᱈ฟ⾡ࡢ᭷ຠᛶࡣⰋࡃ▱ࡽࢀ࡚࠾ࡾ;LH<HWDO .LGQH\,QWࠊᡥ᱈ฟ⭈⓶㉁ ࢫࢸࣟࢻࣃࣝࢫ⒪ἲࢆ᪩ᮇ⾜࠺ࠊ௨ୖࡢᐶ ゎ⋡ࡀᚓࡽࢀࡿࠋ୍᪉ࠊཱྀᡥ᱈࡛⢾ 㙐 ,J$ ࡀ⏘⏕ࡉࢀࡿᮏ㉁ⓗ࡞ᶵᗎࡣᮍゎ࡛᫂࠶ࡿࠋࡋ ࡶࠊ㍍᪩ᮇᑐࡋ࡚くࢆక࠺እ⛉ⓗ⒪ 㔞ࢫࢸࣟࢻ⒪ἲࢆᚲせࡍࡿ࠸࠺ၥ㢟ࡶゎỴࡉ ࢀ࡚࠸࡞࠸ࠋࡑࡢࡓࡵࡇࡢ⒪ࡣࠊᅜእ࡛ࡣࡃㄆ ▱ࡉࢀ࡚࠾ࡽࡎࠊ᭱㏆ࡢ .',*2 ࢞ࢻ࡛ࣛࣥࡣ͆⾜ ࢃ࡞࠸ࡇࢆ᥎ዡ͇ࡉࢀ࡚࠸ࡿࠋࡋࡓࡀࡗ࡚ࠊ ⮫ ᗋ ⓗ࡞᭷ຠᛶࢆ 5&7 ࡛ド᫂ࡍࡿࡶࠊࡑࡢ᭷ຠᛶ ࢆㄝ᫂ࡍࡿ࣓࢝ࢽࢬ࣒⮬యࢆ᫂ࡽࡍࡿࡇࡀᚲ 㡲࡛࠶ࡾࠊࢃࡀᅜࡢࡇࡢศ㔝ᦠࢃࡿ⪅ࡢ㈐ົ࡛ࡶ ࠶ࡿࠋ ⏦ㄳ⪅ࡽࡣᅇࡢ⏦ㄳඛ❧ࡕࠊ,J$ ⭈ࠊ⩦័ᛶ ᡥ᱈⅖ࠊᑠඣᡥ᱈⫧ࡢฟᡥ᱈ࡢ῝㒊㝜❐⤌ ⧊ࡽ '1$ ࢆᢳฟࡋࠊ6U51$9 㡿ᇦࡢࢤࣀ࣒ࢩ࣮ ࢡ࢚ࣥࢫࢆỴᐃࡋࡓࠋ,J$ ⭈࡛ࠊ)XVREDFWHULXP 3UHYRWHOOD7UHSRQHPD࡞ࡀࡓࡿ⣽⳦ᒓࣞ࣋ࣝ ࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ 㸺ᨭࡢෆᐜ㸼 ⌧ᅾࡲ࡛⾜ࢃࢀࡓ 6U51$ ゎᯒ࡛ ,J$ ⭈࡛ࡓ ࡿ ⣽ ⳦ ⩌ ㄆ ࡵ ࡽ ࢀ ࡓ ࠊ )XVREDFWHULXP ᒓ 3UHYRWHOOD ᒓࡘ࠸࡚ࢻࣛࣇࢺࢤࣀ࣒ࢩ࣮ࢡ࢚ࣥࢫ ࢆ⾜࠸ࠊබⓗࢹ࣮ࢱ࣮࣋ࢫ࡞ẚ㍑ࢆ⾜࠺ࠋ,J$ ⭈≉␗ⓗ࡞⣽⳦ᰴࡀᏑᅾࡍࡿࠊࢤࣀ࣒ẚ㍑ゎ ᯒࢆ⾜࡞࠺ࠋ 㸺ᨭࡼࡗ࡚ᮇᚅࡉࢀࡿ◊✲ᡂᯝ⌧≧㸼 ,J$ ⭈ᝈ⪅࡛ࡣ୍ぢṇᖖ࡞ᡥ᱈࡛ࡶฟࡋ࡚ほ ᐹࡍࡿ῝㒊࡛ࡣᕢឤᰁࡀㄆࡵࡽࢀࠊࡲࡓ᎘Ẽᛶ ⳦࡞ࡀ㛵ࡋ࡚࠸ࡿྍ⬟ᛶࡶ࠶ࡾࠊ㏻ᖖࡢᡥ᱈ᇵ 㣴࡛ࡣ᳨ฟ࡛ࡁ࡞࠸ྍ⬟ᛶࡀ࠶ࡿࠋ⥙⨶ⓗ࡞⣽⳦ࢤ ࣀ࣒ゎᯒࡼࡾ ,J$ ⭈ࡢᡥ᱈⤌⧊࠾ࡅࡿ⣽⳦ྀ ࡢㇺࡀ᫂ࡽ࡞ࢀࡤࠊ,J$ ⭈ࡢⓎ࠾ࡅࡿ⣽ ⳦ឤᰁࡢᙺࡀ᫂ࡽ࡞ࡾࠊᡥ᱈⏕᳨ࡼࡿデ᩿ ἲࡢ㛤Ⓨࡸࠊᡥ᱈ฟ⒪ࡢ⌮ㄽⓗ࡞᰿ᣐࡘ࡞ࡀ ࡿᛮࢃࢀࡿࠋ ,J$ ⭈࡛ࡣ⢾㙐 ,J$ ࡢ⏘⏕ࡀஹ㐍ࡋࠊࡉࡽ ࡣ⢾㙐 ,J$ ࣄࣥࢪ㒊ᑐࡍࡿᢠయࡀ⏘⏕ࡉࢀ ࡚ච」ྜయࢆᙧᡂࡋࠊ⭈⣒⌫య࣓ࢧࣥࢠ࣒࢘㡿ᇦ ỿ ╔ ࡋ ࡚ ⣒ ⌫ య ⭈ ⅖ ࡀ ច ㉳ ࡉ ࢀ ࡿ ࠊ PXOWLKLW WKHRU\ࡀᥦၐࡉࢀ࡚࠸ࡿࠋᡥ᱈⣽⳦ࢤࣀ࣒ゎᯒ࡛᫂ ࡽ࡞ࡗࡓ⣽⳦⩌ࡀࡇࢀࡽࡢචᛂࢆច㉳ࡍࡿ ᡥ᱈ᇵ㣴⣽⬊࡛☜ㄆࡍࡿࠋࡲࡓࡇࢀࡽࡢ⣽⳦⩌ࡢ ⳦యᡂศࡀචᢠཎ࡛࠶ࡿྍ⬟ᛶࡶ࠶ࡾࠊせ࡞⣽ ⳦⩌ࡢ⳦యᡂศᑐࡍࡿᢠయࢆసᡂࡋࠊ⾑୰࡛ࡢᢠ య౯ࡢ ᐃࡸࠊ⭈⏕᳨⤌⧊࠾࠸࡚ࡇࢀࡽࡢ⳦యᡂ ศࢆྵࡴච」ྜయࡀᏑᅾࡍࡿ࠺᳨ウࢆ⾜࠺ࠋ ,J$ ⭈≉␗ⓗ࡞⣽⳦⩌ࡣᢠ⳦⸆ࢆ⏝ࡋࡓ᪂ࡓ ࡞⒪ࢱ࣮ࢤࢵࢺ࡞ࡿྍ⬟ᛶࡀࡁ࠸⪃࠼ࡽࢀ ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ᡂᯝࡀࡲࡲࡾḟ➨ࠊⱥᩥㄽᩥࢆᢞ✏ணᐃ - 458 - බເᨭㄢ㢟㸸2014 ᖺᗘ ⏥ࡢゅ㸦ࢶࣀ㸧ᙧᡂ㑇ఏᏊࢿࢵࢺ࣮࣡ࢡࡢ㐍࣓࢝ࢽࢬ࣒ࡢゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ ᪂⨾ ㍤ᖾ࣭ᇶ♏⏕≀Ꮫ◊✲ᡤ㐍Ⓨ⏕◊✲㒊㛛 㸺⫼ᬒ࣭┠ⓗ㸼 ๓⬚㒊ゅࠊ㞤㢌㒊ゅ┦ᙜ㡿ᇦࠊ㞤๓⬚㒊ゅ┦ᙜ㡿ᇦ㸧 ᮏ◊✲ࡣࠊ」ྜ㐺ᛂᙧ㉁ࡋ࡚᪻ࡢከࡃࡢ⣔⤫ ࢆࢧࣥࣉࣝ⏝࠸ࣛࣈ࣮ࣛࣜࢆస〇ࡋࠊ࢜ࣜࢦ࢟ ࡛⊂❧⋓ᚓࡉࢀࡓゅ㸦ࢶࣀ㸧╔┠ࡍࡿࠋ᪻ࢆ ࣕࢵࣉἲᇶ࡙ࡃ 766VHT ゎᯒࢆ⾜ࡗࡓࠋྛࣛࣈ ྲྀࡾᕳࡃᵝࠎ࡞⎔ቃࡢ୰࡛ࠊ⦾Ṫᡓ␎ࡢ┦స⏝ ࣮ࣛࣜࡘ࠸࡚ 0 ࣮ࣜࢻ⛬ᗘ㸦Ⓨ⌧㔞ࡢప࠸㌿ ࢆ㏻ࡋ࡚㐍ࡋࡓ᪂ወ㐺ᛂᙧែ࡛࠶ࡿゅࡣࠊࡢ ᅉᏊࡢ㌿㛤ጞⅬ࡛ࡶㄆ㆑࡛ࡁࡿ⛬ᗘࡢゎᯒつᶍ㸧 ⏕≀⩌ẚ㢮ࡢ࡞࠸ᴟࡵ࡚ከᵝ࡞ᙧែࡀᏑᅾࡍࡿࠋ ࡢ㓄ิሗࢆྲྀᚓࡍࡿࡇࢆ┠ᣦࡋࡓࠋ ◊✲ᮦᩱࡣࠊ㞝ࡢࡳ㢧ⴭⓎ㐩ࡋࡓゅࢆᣢࡘ࢝ ࣈࢺ࣒ࢩࠊࡲࡓ࢝ࣈࢺ࣒ࢩࡣࡑࢀࡒࢀ⊂❧ゅࢆ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⋓ᚓࡋࡓ࢜࢜ࢶࣀࢥࢡࢾࢫࢺࣔࢻ࢟࠾ࡼࡧࣇࣝ +L6HT ࡚ ES6LQJOH(QGࡢ᮲௳࡛ࢩ࣮ࢡ ࢟ࢡ࣒ࢩ࡞ゅᙧ㉁ࢆ᭷ࡍࡿᵝࠎ࡞᪻ࢆ⏝࠸ࡿࠋ ࢚ࣥࢫࢆ⾜ࡗࡓࡇࢁࠊ㞝㢌㒊ゅཎᇶࡣ 0 ࣮ࣜ ࡇࢀࡽ᪻ࡢⅬࡸ⏦ㄳ⪅ࡀࡇࢀࡲ࡛☜❧ࡋࡓ㑇 ࢻࠊ㞝๓⬚㒊ゅཎᇶࡣ 0 ࣮ࣜࢻࠊ㞤㢌㒊ゅ┦ ఏᏊᶵ⬟ゎᯒἲࢆ⏕ࡋࠊḟୡ௦ࢩ࣮ࢣࣥࢧ࣮ ᙜ㡿ᇦཎᇶࡣ 0 ࣮ࣜࢻࠊ㞤๓⬚㒊ゅ┦ᙜ㡿ᇦཎ ࢆ⏝࠸ࡓẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࡼࡾゅᙧ ᇶࡣ 0 ࣮ࣜࢻⰋዲ࡞⤖ᯝࡀᚓࡽࢀࡓࠋ⌧ᅾࠊ ᡂࢆไᚚࡍࡿ㑇ఏᏊࢆྠᐃࡋࠊゅᙧᡂ㑇ఏᏊࢿࢵࢺ ඹྠ◊✲⪅ඹᅇᚓࡽࢀࡓ 766VHT ሗࢆࡇࢀ ࣮࣡ࢡࡢ㐍࣓࢝ࢽࢬ࣒ࢆゎ᫂ࡍࡿࠋ᭱⤊ⓗࡣࠊ ࡲ࡛ゎㄞࡋࡓ࢝ࣈࢺ࣒ࢩࢤࣀ࣒ᴫせ㓄ิᑐࡋ࡚ ✀㺃⛉㺃ୖ⛉࡛ࣞ࣋ࣝ␗࡞ࡿ᪻✀ࢆ⏝࠸ࡓẚ㍑ゎᯒ ࣐ࢵࣆࣥࢢࢆ⾜ࡗ࡚࠸ࡿࠋ ࡼࡾࠊ㐍ࡢ᪂ࡋ࠸ඹ㏻⌮ㄽࡢᵓ⠏ࢆ┠ᣦࡍࠋ ࡇࢀࡲ࡛࢝ࣈࢺ࣒ࢩ࡛ࡣࠊ⥙⨶ⓗ࡞ࢤࣀ࣒ゎᯒ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᢏ⾡ࢆ㥑ࡋ࡚㢧ⴭ࡞ᛶⓗᆺᙧ㉁࡛࠶ࡿゅࡢᙧᡂ ࡇࢀࡲ࡛ࠊ㌿ᅉᏊࢆࢥ࣮ࢻࡍࡿᛶỴᐃ㑇ఏᏊ ࣓࢝ࢽࢬ࣒ࢆゎ᫂ࡍࡿࡓࡵࠊࢤࣀ࣒ゎㄞࡸ 51$VHT GRXEOHVH[GV[ࡣゅᙧᡂᴟࡵ࡚㔜せ࡞ᙺࢆᯝࡓ ἲࢆ⏝࠸ࡓẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ⾜ࡗ࡚ ࡍࡇࢆ᫂ࡽࡋ࡚ࡁࡓࠋᮏᨭࡼࡾᚓࡽࢀࡿ ࡁࡓࠋࢤࣀ࣒ゎㄞࡣࠊVFDIIROG1 ࡀ 0E Ⰻ ࢤࣀ࣒࣡ࢻ࡞㌿㛤ጞⅬሗຍ࠼ࠊ⌧ᅾゎᯒ୰ ዲ࡞⤖ᯝࡀᚓࡽࢀࠊࢤࣀ࣒ࢧࢬࡣ⣙ 0E ࡛࠶ ࡢ 'V[ ࡢࢤࣀ࣒࣡ࢻ࡞⤖ྜ㓄ิሗࢆ⤫ྜࡍࡿࡇ ࡿࡇࡀุ᫂ࡋࡓࠋࡲࡓࠊゅཎᇶࡢ 51$VHT ゎᯒ ࡼࡾࠊGV[ ᶆⓗ㑇ఏᏊࡢࢤࣀ࣒࡛ࣞ࣋ࣝࡢヲ⣽࡞ ࡼࡾࠊᛶ㛫ẚ㍑࠾ࡼࡧᛶෆẚ㍑ࡼࡾⓎ⌧ᕪࡢ࠶ࡿ ྠᐃࡀྍ⬟࡞ࡿࠋࡇࢀࡽᶆⓗ㑇ఏᏊࡢೃ⿵ࡘ࠸ 㑇ఏᏊࢆࣜࢫࢺࢵࣉࡋࠊ51$L ἲࡼࡿᶵ⬟ゎᯒࢫ ࡚ࠊ51$L ἲࡼࡿᶵ⬟ゎᯒࢆ⾜࠸ゅᙧᡂࡢ㛵ࢆ ࢡ࣮ࣜࢽࣥࢢࡼࡾࠊከᩘࡢゅᙧᡂ㑇ఏᏊࡢྠᐃ ᳨ドࡍࡿࠋ௨ୖࡢゎᯒࡼࡾࠊゅᙧᡂ㑇ఏᏊࢿࢵࢺ ᡂຌࡋࡓࠋ ࣮࣡ࢡ࠾ࡅࡿ GV[ ࡢᙺࡢゎ᫂ࢆ┠ᣦࡍࠋ ᮏࢤࣀ࣒ᨭࣉࣟࢪ࢙ࢡࢺ࡛ࡣࠊ࢝ࣈࢺ࣒ࢩ࠾ ࡲࡓࠊ㌿㛤ጞⅬࡢ⥙⨶ⓗྠᐃࡼࡾ㠀ࣔࢹࣝ᪻ ࡅࡿゅᙧᡂ㑇ఏᏊࢿࢵࢺ࣮࣡ࢡࡢㇺゎ᫂ྥࡅࠊ ࠾࠸࡚ࡶ᭷ຠ࡞᪂つ LQYLYR ࢚ࣞࢡࢺ࣏࣮ࣟࣞ ㌿㛤ጞⅬ㏆࠸㌿ᅉᏊ⤖ྜࢧࢺࡢ≉ᐃ᭷ຠ ࢩࣙࣥἲ㸦$QGRDQG)XMLZDUD㸧ࢆ⏝࠸ࡓࣉࣟ ࡞ 766VHT ἲࡼࡾࠊ࢝ࣈࢺ࣒ࢩࡢゅཎᇶ࡛Ⓨ⌧ࡍ ࣮ࣔࢱ࣮㡿ᇦࡢ࣏࣮ࣞࢱ࣮ࢵࢭࢆᐜ᫆⾜࠺ࡇ ࡿ㑇ఏᏊࡘ࠸࡚㌿㛤ጞⅬࡢ⥙⨶ⓗྠᐃࢆヨࡳࡓࠋ ࡀྍ⬟࡞ࡾࠊࡉࡽ࡞ࡿࢤࣀ࣒ᶵ⬟ࡢゎ᫂㈨ࡍ ࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭࡢෆᐜ㸼 766VHT ἲࡼࡿ㌿㛤ጞⅬࡢ⥙⨶ⓗྠᐃ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ୖ㏙ࡋࡓ 51$VHT ゎᯒྠᵝࠊ๓⻈ ᪥ࡢ㔝⏕ ࡞ࡋ㸦ᮏᨭෆᐜ࡛ᚓࡽࢀࡓ⤖ᯝࢆ࢝ࣈࢺ࣒ࢩࡢ ᆺࡢ㞤㞝ࡑࢀࡒࢀࡢྛཎᇶ㸦ヨᩱᩘ 㸹㞝㢌㒊ゅࠊ㞝 ࢤࣀ࣒ゎㄞࡢㄽᩥྵࡵ࡚Ⓨ⾲ࡍࡿணᐃ࡛࠶ࡿࠋ 㸧 - 459 - බເᨭㄢ㢟㸸2014 ᖺᗘࠊ2015 ᖺᗘ ࢸࣥࢺ࣒࢘ࢩ㛵㐃ࡋࡓᨃែᩬ⣠ࢆࡶࡓࡽࡍࢤࣀ࣒࣭㑇ఏᏊ┦㛵ࡢゎ᫂ ᪂Ꮫ⾡㡿ᇦ◊✲ ᪂⨾ ㍤ᖾ࣭ᇶ♏⏕≀Ꮫ◊✲ᡤ㐍Ⓨ⏕◊✲㒊㛛 㸺⫼ᬒ࣭┠ⓗ㸼 ᯒ ᮏ◊✲ࡣࠊࢼ࣑ࢸࣥࢺ࢘ࡢᴟࡵ࡚ከᵝᛶᐩࡴᩬ ⩟ཎᇶࢆ 51$ ᢳฟࡢࢧࣥࣉࣝࡋࠊ⤫ィⓗⓎ⌧ ⣠ከᆺ࠾ࡼࡧࢸࣥࢺ࣒࢘ࢩ㛵㐃ࡋࡓᨃែᩬ⣠╔ 㔞᭷ពᕪࡢ࠶ࡿ㑇ఏᏊࢆࣜࢫࢺࢵࣉࡍࡿࡓࡵࠊ ┠ࡍࡿࠋ◊✲ᮦᩱࡣࠊྛ✀ࢸࣥࢺ࣒࢘ࢩ࠾ࡼࡧࢸ ELRORJLFDOUHSOLFDWH ࡣ ࢧࣥࣉࣝ‽ഛࡋࡓࠋྜィ ࣥࢺ࣒࢘ࢩࡣ⊂❧ᨃែᩬ⣠ࢆ⋓ᚓࡋࡓࢸࣥࢺ࢘ ࡢࣛࣈ࣮ࣛࣜࢆస〇ࡋࠊ+L6HT ࢆ⏝࠸࡚ ࣀ࣑ࣁ࣒ࢩࢆ⏝࠸ࡿࠋ⏦ㄳ⪅ࡀୡ⏺ඛ㥑ࡅ࡚ࢼ࣑ ES3DLUHG(QGࡢ᮲௳࡛ࢩ࣮ࢡ࢚ࣥࢫࢆ⾜ࡗࡓࡇ ࢸࣥࢺ࢘࠾࠸࡚ྠᐃࡋࡓ⩟యࡢᩬ⣠ࣃࢱ࣮ࣥࢆ ࢁࠊྛࢧࣥࣉࣝࡘࡁ ࠥ0UHDG ࡢ㓄ิࢆᚓࡿ Ỵᐃࡍࡿ㌿ᅉᏊࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊ㸦ᩬ⣠ࣉࣞࣃ ࡇࡀ࡛ࡁࡓࠋ ࢱ࣮ࣥ㑇ఏᏊ㸧↔Ⅼࢆ⤠ࡾࠊᮦᩱ⏝࠸ࡿ᪻ࡢ Ⅼࡸ⏦ㄳ⪅ࡀࡇࢀࡲ࡛☜❧ࡋࡓ㑇ఏᏊᶵ⬟ゎᯒ 2015 ᖺᗘ ἲࢆάࡋࠊᩬ⣠ከᆺࢆࡶࡓࡽࡍࢤࣀ࣒ኚࠊࢸࣥ 㸦㸧766VHT ἲࡼࡿ㌿㛤ጞⅬࡢ⥙⨶ⓗྠᐃ ࢺ࣒࢘ࢩ⛉ෆࡢ␗࡞ࡿᩬ⣠ࣃࢱ࣮ࣥࡸ⊂❧⋓ᚓࡉ ⌧ᅾࠊࢧࣥࣉࣝ‽ഛ୰ࠋ ࢀࡓᨃែᩬ⣠ࢆࡶࡓࡽࡍ㑇ఏᏊⓎ⌧ไᚚ㡿ᇦ࡞ࡢ 㸦㸧ࣄ࣓࣓࢝ࣀࢥࢸࣥࢺ࢘ཬࡧࢲࣥࢲࣛࢸࣥࢺ࢘ࡢ ࢤࣀ࣒ኚࠊࡉࡽ」㞧࡞ᑐ❧㑇ఏᏊᵓᡂࡀ⏕ࡌࡓ ࢻࣛࣇࢺࢤࣀ࣒ゎㄞ ࢤࣀ࣒せᅉࢆ≉ᐃࡋࠊᨃែᩬ⣠ࡀࡶࡓࡽࡉࢀࡓࢤࣀ ࢧࣥࣉࣝ㏦῭ࡳࠋ⌧ᅾࠊゎᯒ୰ࠋ ࣒࣭㑇ఏᏊ┦㛵ࢆゎ᫂ࡍࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 㸺ᨭࡢෆᐜ㸼 ๓ᖺᗘࡢࢤࣀ࣒ᨭࡼࡾࢤࣀ࣒ゎㄞࢆ⾜ࡗࡓࢼ ᖺᗘ ࣑ࢸࣥࢺ࢘ࡣࡑࢀࡒࢀ⊂❧ᩬ⣠ከᆺࢆ⋓ᚓࡋࡓ 㸦㸧ࢼ࣑ࢸࣥࢺ࢘ࡢࢻࣛࣇࢺࢤࣀ࣒ゎㄞ ࣄ࣓࣓࢝ࣀࢥࢸࣥࢺ࢘ཬࡧࢲࣥࢲࣛࢸࣥࢺ࢘╔┠ 㸦㸧51$VHT ࢆ⏝࠸ࡓẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎ ࡋࡓࠋࢼ࣑ࢸࣥࢺ࢘ຍ࠼ࠊᮏࢤࣀ࣒ᨭ࠾࠸࡚ࠊ ᯒ ⊂❧ࡋࡓᩬ⣠ከᆺࢆࡶࡘࣄ࣓࣓࢝ࣀࢥࢸࣥࢺ࢘ཬࡧ ࢲࣥࢲࣛࢸࣥࢺ࢘ࡢࢤࣀ࣒ゎㄞࢆ⾜࠺ࡇࡼࡾࠊ ᖺᗘ 㢧ⴭ࡞ᩬ⣠ከᆺࢆ᭷ࡍࡿ 3 ✀ࡢࢸࣥࢺ࣒࢘ࢩࡢẚ㍑ 㸦㸧766VHT ἲࡼࡿ㌿㛤ጞⅬࡢ⥙⨶ⓗྠᐃ ࢤࣀ࣒ゎᯒࡀྍ⬟࡞ࡿࠋ᭱㏆ࠊ⏦ㄳ⪅ࡣ RNAi ἲ 㸦㸧ࣄ࣓࣓࢝ࣀࢥࢸࣥࢺ࢘ཬࡧࢲࣥࢲࣛࢸࣥࢺ࢘ࡢ ࢆ㥑ࡍࡿࡇࡼࡾࠊࣄ࣓࣓࢝ࣀࢥࢸࣥࢺ࢘࠾ ࢻࣛࣇࢺࢤࣀ࣒ゎㄞ ࠸࡚ࢼ࣑ࢸࣥࢺ࢘ࡣ␗࡞ࡿᩬ⣠ࣉࣞࣃࢱ࣮ࣥ㑇ఏ Ꮚࡢྠᐃᡂຌࡋࡓࠋᮏ◊✲࡛ࡣࠊࡇࢀࡲ࡛ྠᐃ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡋࡓᩬ⣠ࣉࣞࣃࢱ࣮ࣥ㑇ఏᏊ↔Ⅼࢆ⤠ࡾࠊ3 ✀㛫࡛ ᖺᗘ ࢤࣀ࣒ᵓ㐀ࡢẚ㍑ཬࡧࡇࢀࡲ࡛☜❧ࡋࡓ✀ࠎࡢ㑇 㸦㸧ࢼ࣑ࢸࣥࢺ࢘ࡢࢻࣛࣇࢺࢤࣀ࣒ゎㄞ ఏᏊᶵ⬟ゎᯒࢆ㏻ࡋ࡚ࠊᩬ⣠ከᆺࢆ⏕ࡌࡉࡏࡿ」㞧 ࢧࣥࣉࣝࡣ㞤ᡂࢆ⏝࠸ࡓࠋ3ODWDQXV ࢆ⏝࠸ࡓ ࡞ᑐ❧㑇ఏᏊᵓᡂࢆࡶࡓࡽࡍࢤࣀ࣒せᅉࢆ≉ᐃࡍࡿ ࢭࣥࣈࣝࡢ⤖ᯝࠊ6FDIIROG1 ࡀ 0E 㠀ᖖ ࡇࢆ┠ᣦࡍࠋ Ⰻዲ࡞⤖ᯝࡀᚓࡽࢀࡓࠋࡲࡓࠊࢤࣀ࣒ࢧࢬࡶ᫂ࡽ ࡞ࡗࡓࠋḟࠊ㑇ఏᏊᵓ㐀ண ࢆ⾜ࡗࡓࠋᚋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡣࠊᅇᚓࡽࢀࡓࢤࣀ࣒ሗࢆ⏝ࡋ࡚ࠊᩬ⣠ከᆺ ࡞ࡋ㸦⌧ᅾゎᯒ୰࡛࠶ࡾࠊゎᯒ⤖ᯝࡀᚓࡽࢀࡓẁ ࢆࡶࡓࡽࡍࢤࣀ࣒ୖࡢせᅉࢆ≉ᐃࡍࡿࠋ 㝵࡛ㄽᩥࡢෆᐜࢆ᳨ウࡍࡿࠋ㸧 㸦㸧51$VHT ࢆ⏝࠸ࡓẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎ - 460 - බເᨭㄢ㢟㸸2015 ᖺᗘ ᪻⩟ࡢ㉳※ከᵝ࣓࢝ࢽࢬ࣒ࡢゎ᫂ࡑࡢᛂ⏝ ᇶ┙◊✲㸦B㸧 ᪂⨾ ㍤ᖾ࣭ᇶ♏⏕≀Ꮫ◊✲ᡤ㐍Ⓨ⏕◊✲㒊㛛 㸺⫼ᬒ࣭┠ⓗ㸼 ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ 㣕⩧ჾᐁࡋ࡚⋓ᚓࡉࢀࡓ᪻ࡢ⩟ࡣࠊ᪻ࡢከ ࢧࣥࣉࣝ㏦῭ࡳࠋ⌧ᅾࠊゎᯒ୰ࠋ ᵝక࠸ࡑࢀࡒࢀࡢ᪻┠࠾࠸࡚≉ᚩࡢ࠶ࡿ⩟ 㸦㸧51$L ฎ⌮ಶయࡢ 51$VHT ࢆ⏝࠸ࡓẚ㍑ࢺࣛࣥࢫ 㐍ࡋ࡚ࡁࡓࠋ⏥㸦㠧⩟┠᪻㸧ࡢሙྜࠊయ ࢡࣜࣉࢺ࣮࣒ゎᯒ ࡢಖㆤᶵᵓࢆᯝࡓࡍࡓࡵ๓⩟ࡀ◳ࡋࡓ㠧⩟ኚ ࢧࣥࣉࣝ‽ഛ୰ࠋ ࡋࠊᚋ⩟ࡣ㣕⩧ჾᐁࡋ࡚ࡢᙺࢆ⥔ᣢࡋ࡚࠸ࡿࠋ 37 ✀࠸࠺ᅽಽⓗ࡞✀ᩘࢆࡿ⏥㸦㠧⩟┠᪻㸧 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢆ≉ᚩࡅࡿᙧ㉁ࡣ๓⩟ࡀ◳ࡋࡓ㠧⩟࡛࠶ࡿࡀࠊ ᮏࢤࣀ࣒ᨭ࡛ࡣࠊṇᖖⓎ⏕㐣⛬ࡢ⩟ཎᇶࡢẚ㍑ ࡑࡢ㠧⩟ᙧᡂ࣓࢝ࢽࢬ࣒ࡢゎ᫂ࡣ࠶ࡲࡾ㐍ࢇ࡛࠸࡞ ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ࠾ࡼࡧ㠧⩟ᙧᡂࡢ㘽࡞ ࠸ࠋ࡛᪻ࡣ୍⯡ࠊ୰⬚Ꮡᅾࡍࡿ๓⩟ࡀࢹࣇ࢛ ࡿ㌿ᅉᏊࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࡢ RNAi ฎ⌮༊ᑐ ࣝࢺࡢᙧែࡉࢀࠊᚋ⬚ࢆ≉ᚩࡅࡿ࣓࣍࢜ࢸࢵ ↷༊ࡢẚ㍑ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆ⾜࠺ࡇ ࢡ㑇ఏᏊ࡛࠶ࡿ Ultrabithorax㸦Ubx㸧ࡼࡾᚋ⩟ ࡼࡾࠊࡇࢀࡲ࡛࡛᫂࠶ࡿ㠧⩟ᙧᡂ㑇ఏᏊ⩌࡞ࡽࡧ ኚࡀࡶࡓࡽࡉࢀ࡚࠸ࡿࠋࡇࢀᑐࡋࠊ㠧⩟┠᪻ 㠧⩟ᙧᡂࡢ㘽࡞ࡿ㌿ᅉᏊࡢᶆⓗ㑇ఏᏊ⩌ࡢೃ ࡛ࡣ⯆῝࠸ࡇࠊࡢ᪻ࡣ㏫ࡢ⌧㇟ࡀほᐹ ⿵ࢆ⥙⨶ⓗࣜࢫࢺࢵࣉࡍࡿࠋḟࠊᖺᗘࡢࢤ ࡉࢀࡿࠋࡘࡲࡾࠊUbx ࡢ RNAi ࡼࡾ㣕⩧ࡢࡓࡵࡢ ࣀ࣒ᨭࡼࡾゎㄞࡋࡓࢼ࣑ࢸࣥࢺ࢘ࡢࢤࣀ࣒ሗ ᚋ⩟ࡀ๓⩟ࡢ≉ᚩࢆᣢࡘ㠧⩟ኚࡍࡿࡢ࡛࠶ࡿࠋ ࢆ⏝ࡋ࡚ࠊࡇࢀࡽೃ⿵࡞ࡿᶆⓗ㑇ఏᏊࡢࣉࣟࣔ ࡇࡢ⌧㇟ࡣ㠧⩟ࡢ㐍࠾ࡅࡿࡁ࡞ㅦ࡞ࡗ࡚࠸ ࣮ࢱ࣮ୖὶᇦ࡞Ⓨ⌧ไᚚ㛵ࡍࡿྍ⬟ᛶࡢ࠶ࡿ ࡿࠋ 㡿ᇦࡘ࠸࡚㠧⩟ᙧᡂࡢ㘽࡞ࡿ㌿ᅉᏊࡢ⤖ྜࢥ ᮏࢤࣀ࣒ᨭ࡛ࡣࠊࡇࡢ㐍ୖࡢㅦࢆゎࡃࡓࡵࠊ 㸦1㸧 ࣥࢭࣥࢧࢫ㓄ิࡢ᭷↓ࢆ᳨⣴ࡋ࡚ࠊ┤᥋ࡢᶆⓗ㑇ఏ ṇᖖⓎ⏕㐣⛬ࡢ⩟ཎᇶࡢ RNA-seq ࢆ⏝࠸ࡓẚ㍑ࢺ Ꮚ࡛࠶ࡿྍ⬟ᛶࢆ᳨ウࡍࡿࠋ⥆࠸࡚ࠊ◊✲௦⾲⪅ࡀ ࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒཬࡧ㸦2㸧◊✲௦⾲⪅ࡀ᪤ ☜❧ࡋࡓ larval RNAi ἲࢆ⏝࠸࡚ᶵ⬟ゎᯒࢆ⾜࠺ࡇ ☜❧ࡋࡓ RNAi ἲࢆ⏝ࡋ࡚㠧⩟ᙧᡂࡢ㘽࡞ࡿ㌿ ࡼࡾࠊ㠧⩟ᙧᡂ㐣⛬࠾ࡅࡿྛ㑇ఏᏊࡢᙺࢆ ᅉᏊࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࡢᶵ⬟㜼ᐖࡼࡗ࡚Ⓨ⌧ ᫂ࡽࡍࡿࡇࡼࡾࠊ㠧⩟ᙧᡂࡢㇺゎ᫂㏕ ࡀኚࡍࡿ㑇ఏᏊࡢ⥙⨶ⓗ࡞ྠᐃࢆ┠ᣦࡍࠋᐇ㦂ᮦ ࡿࠋࡉࡽࠊ㠧⩟ᙧᡂ㐣⛬࠾࠸࡚㔜せ࡞ᙺࢆᯝ ᩱࡣࠊᖺᗘࡢࢤࣀ࣒ᨭࡼࡾࢤࣀ࣒ᴫせ㓄ิ ࡓࡍࡇࡀุ᫂ࡋࡓ㑇ఏᏊࡘ࠸࡚ࡣࠊࢩࣙ࢘ࢪࣙ ࢆỴᐃࡋࡓࢼ࣑ࢸࣥࢺ࢘ࢆ⏝࠸ࡿࠋ ࢘ࣂ࢚ࡢ㣕⩧⏝ࡢ๓⩟ࡢⓎ⏕㐣⛬࠾࠸࡚␗ᡤⓗ Ⓨ⌧ࡉࡏࡿࡇࡼࡾࠊ㣕⩧⏝ࡢ⩟ࡽ㠧⩟ᙧ 㸺ᨭࡢෆᐜ㸼 ែⓗ࡞ኚࢆᘬࡁ㉳ࡇࡍᶵ⬟ࢆ᭷ࡍࡿྰࡘ࠸ 㸦㸧ṇᖖⓎ⏕㐣⛬ࡢ⩟ཎᇶࡢ 51$VHT ࢆ⏝࠸ࡓẚ㍑ ࡚ࡶ᳨ドࡍࡿࠋ௨ୖࡢࡼ࠺࡞ loss-of-function ࠾ࡼࡧ ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ gain-of-function ᐇ㦂ࢆ㏻ࡋ࡚㠧⩟┠᪻ࡢ㐍 㸦㸧51$L ฎ⌮ಶయࡢ 51$VHT ࢆ⏝࠸ࡓẚ㍑ࢺࣛࣥࢫ క࠸⏕ࡌࡓ㠧⩟ࡢ㐍ࣉࣟࢭࢫࡢ᥎ᐃࢆ⾜࠺ࠋ ࢡࣜࣉࢺ࣮࣒ゎᯒ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࡞ࡋ㸦⤖ᯝࡀᚓࡽࢀࡓẁ㝵࡛ㄽᩥࡢෆᐜࢆ᳨ウࡍ 㸦㸧ṇᖖⓎ⏕㐣⛬ࡢ⩟ཎᇶࡢ 51$VHT ࢆ⏝࠸ࡓẚ㍑ ࡿ㸧 ࠋ - 461 - බເᨭㄢ㢟㸸2013~2014 ᖺᗘ ᪂つᙧ㉁⋓ᚓࡢࢤࣀ࣒ᇶ┙ࡑࡢ㐍ⓗ㉳※ ᪂Ꮫ⾡㡿ᇦ◊✲ 㝵ᇽ 㞞ே࣭ᮾிᕤᴗᏛᏛ㝔⏕⌮ᕤᏛ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ࠕᖹ⾜㐍ࠖࡣࠊ⣔⤫ⓗ␗࡞ࡿࢢ࣮ࣝࣉࡢ⏕≀ ࡀࠊఝࡓ⏕ᜥ⎔ቃ㐺ᛂࡍࡿࡓࡵࡑࢀࡒࢀࡀ⊂❧ ఝ㏻ࡗࡓᙧែᙧ㉁ࢆ⋓ᚓࡍࡿࡇ࡛࠶ࡿࠋࡇࢀࡣ ᪂つᙧ㉁ࡢ⋓ᚓ࠾࠸࡚⮬↛㑅ᢥࡀ㔜せ࡛࠶ࡿࡇ ࢆⰍ⃰ࡃ♧၀ࡋࠊᙧែ㐍ࡢ࣓࢝ࢽࢬ࣒ࢆ⌮ゎࡍࡿ ୖ࡛㔜せ࡞⌧㇟࡛࠶ࡿࠋᮾࣇࣜ࢝ࡢᆅ⁁ᖏ ⨨ࡍࡿ୕†ࠊࣅࢡࢺࣜ†ࠊ࣐ࣛ࢘†ࠊࢱࣥ࢞ ࢽ࢝†ࡣࠊࡑࢀࡒࢀᅛ᭷࡞✀ࡀᩘⓒࢆ㉸࠼࡚ ⏕ᜥࡋ࡚࠸ࡿࡀࠊࡇࢀࡽࡢࢩࢡࣜࢵࢻࡣ†ࢆ㉸࠼࡚ ఝࡓᙧ㉁ࢆࡶࡘࡇࡀ▱ࡽࢀ࡚࠾ࡾࠊୖ㏙ࡓᖹ ⾜㐍ࡢዲ࠸࠼ࡿࠋᮏ◊✲࡛ࡣࠊࡢ୕† ࠾ࡅࡿࢩࢡࣜࢵࢻ࡛ࡶ࡞ࡽࡎほᐹࡉࢀࡿࠕ၁ࡢ⫧ ࠖ࠸࠺ᖹ⾜㐍ᙧ㉁╔┠ࡋࠊࡑࡢ⋓ᚓ࣓࢝ ࢽࢬ࣒ࢆ᫂ࡽࡋࡼ࠺⪃࠼࡚࠸ࡿࠋලయⓗࡣࠊ ࢩࢡࣜࢵࢻ♽ඛ㞟ᅋࡀ࠸࡞ࡿ㑇ఏⓗᇶ┙㸦ኚ␗ࢧ ࢺ㸧ࢆࡶࡕࠊࡑࢀᑐࡋ࡚ࡢࡼ࠺࡞⮬↛㑅ᢥࡀ ാࡃࡇ࡛ࠕ၁ࡢ⫧ࠖ࠸࠺ᖹ⾜㐍ࡀ㉳ࡁࡓ ࡢࢆ᫂ࡽࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ၁ ࡢ ⫧ ࡋ ࡚ ࠸ ࡿ ࢩ ࢡࣜ ࢵ ࢻ ✀ 㸦 Haplochromis chilotes㸧⫧ࡋ࡚࠸࡞࠸✀㸦H. sauvagei㸧ࡢ 2 ✀ࢆᑐ㇟ࠊࢤࣀ࣒㓄ิࡢỴᐃ㸦ࣜࢩ࣮ࢣࣥࢫ㸧ࠊ ␗✀㛫㞧✀⣔⤫ࡢᵓ⠏ࡽ㑇ఏᏊᆅᅗࡢస〇ࠊQTL ࣐ࢵࣆࣥࢢࢆ࠾ࡇ࡞ࡗ࡚ࡁࡓࠋᮏࢤࣀ࣒ᨭ࠾࠸ ࡚ࡶࠊࡇࢀࡲ࡛ࡢ◊✲ࡢὶࢀࢆࡃࢇ࡛ୖグ 2 ✀ࡘ ࠸࡚ࡢゎᯒࢆ౫㢗ࡋࡓࠋࡲࡎ 2013 ᖺᗘࡣࠊ2 ✀ࡢ ၁⤌⧊ࡘ࠸࡚Ⳣ㔝࣭㕥ᮌ GP RNA-seq ࢆᐇࡋ ࡚࠸ࡓࡔ࠸ࡓࠋࡑࡢ┠ⓗࡣࠊ2 ✀㛫ࡢ၁⤌⧊࠾ࡅࡿ ᵝࠎ࡞㑇ఏᏊⓎ⌧ࡢ⤯ᑐ㔞ࢆẚ㍑ࡍࡿࡇ࡛ࠊ၁ࡢ ⫧㛵ࡍࡿ㑇ఏᏊࢆ᥈⣴ࡍࡿࡇ࠶ࡿࠋヨ ᩱᩘ㛵ࡋ࡚ࡣࠊH. chilotes ၁⤌⧊ RNA x 4 ಶయศ ࠊH. sauvagei ၁⤌⧊ RNA x 4 ಶయศ࡛ࠊࡑࢀࡒࢀ ಶయẖ Tag ᶆ㆑ࡋ Hiseq2000 ࢆ⏝࠸࡚ 2 ࣮ࣞࣥศ ࡢ㓄ิࢹ࣮ࢱࢆᚓࡓࠋࡇࡢᨭࡼࡿゎᯒࡼࡗ࡚ ┠ⓗ㑇ఏᏊࡢ⤠㎸ࡳᡂຌࡋࡓࡓࡵࠊ2014 ᖺᗘࡣࢩ ࢫኚ␗㡿ᇦࡢ≉ᐃࢆࡍࡿࡇࡀ┠ⓗ࡞ࡗࡓࠋࡑࡇ ࡛ୖグ 2 ✀ຍ࠼࡚ࠊ࢘ࢺࢢ࣮ࣝࣉ࡞ࡿ✀㸦L. rufus㸧ࡢ㔝⏕ಶయࢆྛ 6 ಶయࡘ࠸࡚ࠊࢤࣀ࣒㓄 ิࢹ࣮ࢱࡢ㞟㸦ྛ✀ 1 ࣮ࣞࣥ㸧ࢆࡋ࡚࠸ࡓࡔ࠸ࡓࠋ ࢭࣥࣈࣝసᴗࡣᮾᕤ࣭ఀ⸨ GP ᐇࡋ࡚࠸ࡓࡔ ࠸ࡓࠋゎᯒࢆ㐍ࡵࡿ࠶ࡓࡾ┠ⓗ㑇ఏᏊࡢ࿘㎶㡿ᇦ ࡀከᆺᐩࢇ࡛࠾ࡾࠊṇ☜࡞ࢭࣥࣈࣝࢆ࠾ࡇ࡞࠺ ࡓࡵ㏣ຍࡢࢩ࣮ࢣࣥࢫࡶᐇࡋ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࣅࢡࢺࣜ†⏘ࢩࢡࣜࢵࢻ H. chilotes㸦၁ཌ㸧 H. sauvagei㸦၁ⷧ㸧ࡢ␗✀㛫㞧✀⣔⤫ࡢ QTL ゎᯒࡢ⤖ ᯝࠊ㐃㙐⩌ 22 ࠾࠸࡚ LOD ್ࡀ 10 ࢆ㉸࠼ࡿ㡿ᇦࡀ ᳨ฟࡉࢀࠊࡇࡢ࣐࣮࣮࢝㏆ഐ MAGP4 㑇ఏᏊ⩌㸦ࢤ ࣀ࣒ゎᯒࡼࢀࡤ 7 ࢥࣆ࣮㸧ࡀᏑᅾࡍࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋ2013 ᖺᐇࡋࡓ RNAseq ࢹ࣮ࢱࢆ Broad Institute ࡼࡗ࡚Ỵᐃࡉࢀࡓࣅࢡࢺࣜ†⏘ࢩ ࢡࣜࢵࢻ Pundamilia nyererei ࡢࢻࣛࣇࢺࢤࣀ࣒㓄ิࠊ ࠾ࡼࡧ MAGP4 㑇ఏᏊ㡿ᇦࢆྵࡴ H. chilotes ࡢ BAC clones ࡢ㓄ิ㸦4 clones㸧ᑐࡋ࡚࣐ࢵࣆࣥࢢࡍࡿࡇ ࡛ MAGP4 㑇ఏᏊ⩌ࡢⓎ⌧ࢆ✀㛫ẚ㍑ࡋࡓࠋࡑࡢ ⤖ ᯝ ࠊ 7 ࢥ ࣆ ࣮ ࠶ ࡿ MAGP4 㑇 ఏ Ꮚ ࡢ ࠺ ࡕ ࠊ MAGP4-CD 㑇ఏᏊࡢࡳࡀࠊH. chilotes ࡢ၁⤌⧊࠾ ࠸࡚㢧ⴭⓎ⌧㔞ࡀప࠸ࡇࡀ᫂ࡽ࡞ࡾࠊࡇࢀ ࡣ qPCR ࡼࡗ࡚ࡶ☜ㄆࡉࢀࡓࠋMAGP4 ࡣᙎᛶ⧄⥔ 㸦࢚ࣛࢫࢳࣥ㸧ࡢᙧᡂ㛵ࡋࠊࡇࡢ㑇ఏᏊࢆྵࡴ ࢤ ࣀ ࣒ 㡿 ᇦ ࡢ Ḟ ᦆ ࡣ ࣄ ࢺ 㑇 ఏ 㸦 Smith-Magenis Syndrome㸧ࢆᘬࡁ㉳ࡇࡍࡇࡀ▱ࡽࢀ࡚࠸ࡿࡀࠊࡑ ࡢ SMS ࡢ≉ᚩࡢ 1 ࡘࡀ၁ࡢ⫧ཌ࡛࠶ࡿࡇࡀグ㍕ࡉ ࢀ࡚࠸ࡿࠋ⚾ࡣ H. chilotes ࠾ࡅࡿ၁ࡢ⫧ࡀ MAGP4 㑇ఏᏊࡢⓎ⌧పୗ㉳ᅉࡍࡿ࢚ࣛࢫࢳࣥ⧄ ⥔ࡢᙧᡂࡼࡿࡶࡢ࡛࠶ࡿணࡋࡓࠋᐇ㝿ࠊ ࢚ࣛࢫࢳ࢝࣡ࣥࢠ࣮ࢯࣥᰁⰍἲࢆ⏝࠸࡚ࢩࢡࣜࢵࢻ ࡢ၁┿⓶࠾ࡅࡿ࢚ࣛࢫࢳࣥ⧄⥔ࡢᵓ㐀ࢆ✀㛫ẚ㍑ ࡋࡓࡇࢁࠊH. chilotes ࠾࠸࡚ࠊ᫂ࡽ࢚ࣛࢫࢳ ࣥ⧄⥔ࡀῶᑡࡋ࡚࠸ࡿࡇࡶ᫂ࡽ࡞ࡗࡓࠋ⚾ࡣࠊ ࡇࡢ MAGP4-CD 㑇ఏᏊࡢⓎ⌧㔞పୗࡀ cis 㡿ᇦࡢ ኚ␗ࡼࡿࡶࡢணࡋࠊࡑࡢኚ␗ࡢ≉ᐃࢆ┠ᣦࡋ ࡚ࣅࢡࢺࣜ†⏘ࢩࢡࣜࢵࢻ 3 ✀ࡢ㔝⏕ಶయྛ 6 ಶ యࡘ࠸࡚ࢤࣀ࣒㓄ิࡢỴᐃࢆࡋ࡚࠸ࡓࡔ࠸ࡓࠋ ゎᯒࡢ⤖ᯝࠊMAGP4-CD 㡿ᇦࡣ㓄ิࡢᤄධࡸ㔜 」ࡀከࡃࠊゎᯒ⏝࠸ࡿ࣮ࣜࢻᩘࢆቑࡸࡍᚲせᛶࡀ ฟ࡚ࡁࡓࡓࡵࠊ⌧ᅾࡣࡑࡢసᴗࢆࡋ࡚㡬࠸࡚࠸ࡿ ࡇࢁ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡇࢀࡲ࡛ᡃࠎࡢࢢ࣮ࣝࣉ࠾ࡅࡿ㐍ゎᯒ◊✲࡛ࡣ ೃ⿵㑇ఏᏊࢆணࡋ࡚ಶู㑇ఏᏊ㓄ิẚ㍑ࢆ࠾ࡇ ࡞ࡗ࡚࠸ࡓࡀࠊᮏᨭࡼࡿ⥙⨶ゎᯒࡼࡗ࡚ࠊࡲ ࡗࡓࡃணࡶࡋ࡚࠸࡞ࡗࡓ㑇ఏᏊ㸦MAGP4 㑇ఏᏊ㸧 ࡢⓎ⌧㔞ኚࢆ᳨ฟࡍࡿࡇࡀ࡛ࡁࡓࡢࡣᡂᯝࡋ ࡚㔜せ࡛࠶ࡗࡓࠋQTL ࣐ࢵࣆࣥࢢࢆࡋ࡚ࡶࠊࡑࢀ௨ 㝆ࡢ㑇ఏᏊ᥈⣴ࡀ㞴ࡋ࠸ࡢࡀᖖ࡛࠶ࡗࡓࡀࠊᅇࡣ RNAseq ࢆᐇࡋ࡚࠸ࡓࡔ࠸ࡓࡼࡗ࡚ࠊࡁ࡞◊ ✲ୖࡢ㐍ᒎࡀ࠶ࡗࡓࠋࡑࡋ࡚ࠊࡇࡢ cis 㡿ᇦࢆ†ࡢ ࢩࢡࣜࢵࢻࢆྵࡵ࡚✀㛫㓄ิẚ㍑ࡍࡿࡇࡸ⮬↛㑅 ᢥࡢ᭷↓ࢆ᳨ฟࡍࡿࡇ࡛ࠊ၁ࡢ⫧㛵ࢃࡿᖹ ⾜㐍ࡢ࣓࢝ࢽࢬ࣒ࢆ᫂ࡽࡋ࡚࠸ࡁࡓ࠸ࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Kudo, Y. et al., GENE 560, 156-164 (2015) Brawand, D. et al., Nature 513, 375-381 (2014) Nikaido, M. et al., Genome Biol. Evol. 6, 1135-1144 (2014) - 462 - බເᨭㄢ㢟㸸2015 ᖺᗘ ᗈ⠊࡞⬨᳝ື≀ඹ㏻ࡢ᪂つࣇ࢙ࣟࣔࣥཷᐜయࡢゎᯒ ᇶ┙◊✲㸦㹁㸧 㝵ᇽ 㞞ே࣭ᮾிᕤᴗᏛᏛ㝔⏕⌮ᕤᏛ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ⌧ᅾࡢᆅ⌫ୖࡣᐇከᵝ࡞⏕≀ࡀᏑᅾࡋ࡚࠸ࡿࡀࠊ ࡇࡢⳘ࡞✀ከᵝᛶࡣ⏕≀ࡢࡶࡘᏛឤぬࡼࡿ✀ ෆ࣭✀㛫࡛ࣞ࣋ࣝࡢㄆ▱ࡼࡗ࡚ᕦጁ⥔ᣢࡉࢀ࡚ ࡁࡓ⚄⛎ࡢ⏘≀ࡶ࠸࠼ࡿࠋࡑࡢ୰࡛ࡶ≉㧗➼⬨ ᳝ື≀ࡗ࡚ࠊႥぬࡸࣇ࢙ࣟࣔࣥឤぬࡣ㠀ᖖྂ ࡃࡽᏑᅾࡍࡿཎጞឤぬ࡛࠶ࡾࠊ⏕Ṫࡸྠ✀ㄆ▱࡞ ࠊ⏕άືࡶࡗࡶᇶᮏⓗ࡞⾜ືྍḞ࡛࠶ ࡿࡇࡀศࡗ࡚࠸ࡿࠋᅇࠊᡃࠎࡣ࣏ࣜࣉࢸࣝࢫ ࡸࢩ࣮ࣛ࢝ࣥࢫ࡞ࡢྂ௦㨶ࡽ㝣⏕ࡢ့ங㢮ࡰ ࡚Ώࡿᗈ⠊࡞⬨᳝ື≀ࢤࣀ࣒୰ඹ㏻ࡢࣇ࢙ࣟ ࣔࣥཷᐜయ㑇ఏᏊ㸦᪂つⓎぢࡉࢀࡓᮍྠᐃ㑇ఏᏊ ࡛ ancV1R ྡࡅࡓ㸧ࡀᏑᅾࡍࡿࡇࢆ᫂ࡽࡋ ࡓ㸦ᅗ㸯࣭⾲㸯ཧ↷㸧 ࠋࡇࡢࡇࡣࠊ⬨᳝ື≀ࡢ㐍 ࡢṔྐࡢ࡞࡛ࠊỈ⏕ࡽ㝣⏕ࡢࢲࢼ࣑ࢵࢡ࡞ ⎔ቃኚࡀ࠶ࡗࡓࡶ㛵ࢃࡽࡎࠊ5 ൨ᖺ௨ୖࡶࡢ㛫ࠊ ඹ㏻ࡢࣇ࢙ࣟࣔࣥཷᐜࢩࢫࢸ࣒ࡀࡎࡗ⥔ᣢࡉࢀ࡚ ࡁࡓࡇࠊࡉࡽࡣ㨶ࡽ့ங㢮ࡲ࡛ඹ㏻ࡢ⏕Ṫࢩ ࢫࢸ࣒ࡀᏑᅾࡍࡿࡇࢆ♧၀ࡍࡿࠋࡇࢀࡣࠊ㐍Ꮫ࣭ ⏕⌮Ꮫⓗࡶ㠀ᖖ㦫ࡃࡁⓎぢ࡛࠶ࡿࠋᮏ◊✲࡛ ࡣࠊࡇࡢ᪂つࣇ࢙ࣟࣔࣥཷᐜయ㑇ఏᏊࡢᶵ⬟ࢆᗈ⠊ ࡞⬨᳝ື≀ࡘ࠸࡚ㄪࠊ࠸ࡲࡔᏑᅾࡀ▱ࡽࢀ࡚ ࠸࡞࠸㨶ࡽ့ங㢮ࡲ࡛ඹ㏻ࡢࣇ࢙ࣟࣔࣥཷᐜࢩࢫ ࢸ࣒ࡢᐇែࢆ᫂ࡽࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᮏ◊✲ࢆ㐍ࡵࡿ࠶ࡓࡾணእ࡞Ⓨぢࡋ࡚ࠊ◳㦵 㨶ࡢ୰࡛ࡶ┿㦵㨶㢮ࡣ ancV1R ࡀᏑᅾࡋ࡞࠸ࡀࠊࡇ ࢀࡲ࡛ࢤࣀ࣒㓄ิࡀබ㛤ࡉࢀ࡚࠸ࡿ࠸ࡃࡘࡢୗ ᮲㫅㢮㸦࠸࠺ྂ௦㨶㸧ࡢࢢ࣮ࣝࣉࡣ ancV1R ࡀ Ꮡᅾࡍࡿࡇ࡛࠶ࡿ㸦⾲㸯ཧ↷㸧ࠋ㐍ⓗ࡞ࢱ࣑ࣥ ࢢࢆ⪃៖ࡍࡿࠊancV1R ࡀ㌾㦵㨶㢮ࡢศᒱᚋㄌ⏕ ࡋࡓᚋࠊከࡃࡢ⬨᳝ື≀࡛ࡣ㧗ᗘಖᏑࡉࢀࡘࡘࡶࠊ ┿㦵㨶㢮࠾࠸࡚ࡣഇ㑇ఏᏊ㸦ᾘኻ㸧ࡋࡓ⪃࠼ ࡽࢀࡿࠋ┿㦵㨶㢮ࡢႥୖ⓶࠾࠸࡚ࡣࠊࣇ࢙ࣟࣔࣥ ཷᐜ⣽⬊㸦㗟㰯⚄⤒⣽⬊㸧ໝ࠸ཷᐜ⣽⬊㸦Ⴅ⚄⤒ ⣽⬊㸧ࡀჾᐁࡋ࡚༊⏬ࡉࢀ࡚࠸࡞࠸ࡇࡀ▱ࡽ ࢀ࡚࠸ࡿࡇࡽࠊࡇࡢ ancV1R ࡀࡇࢀࡽࡢ⚄⤒⣽⬊ ࡢ༊⏬㸦ࡘࡲࡾ㗟㰯ჾᐁࡢᙧᡂ㸧㛵ࡋ࡚࠸ࡿ ྍ⬟ᛶࡶ࠶ࡾ⯆῝࠸ࠋࡓࡔࡋࠊୗ᮲㫅㨶㢮┿ 㦵㨶㢮ࡢ⣔⤫ⓗ࡞㛵ಀᛶࡣ᭕࡛࠶ࡾࠊ≉ཎጞⓗ ࡞┿㦵㨶㢮࡛࠶ࡿࣟ࣡ࢼ㢮㛵ࡋ࡚ࡣࠊࡲࡔ ancV1R ࡀᏑᅾࡍࡿྍ⬟ᛶࡶ࠶ࡿࠋࡲࡓࠊୗ᮲㫅㢮 ࡸཎጞⓗ࡞┿㦵㨶㢮࠾ࡅࡿႥୖ⓶ࡢヲ⣽࡞ᵓᡂ 㸦༊⏬ࡀ࠶ࡿྰ㸧ࡣ᫂ࡽ࡞ࡗ࡚࠾ࡽࡎࠊ ⬨᳝ື≀࠾ࡅࡿ ancV1R 㗟㰯ჾᐁᙧᡂࡢ㛵㐃ᛶ ࢆ᫂ࡽࡍࡿୖ࡛ࡣࠊୖグࡢ㨶✀ࡢႥୖ⓶࠾ࡅ ࡿ ancV1R ࡢ᭷↓ࡸࠊႥぬཷᐜయ㑇ఏᏊࡢⓎ⌧ࣃࢱ࣮ ࣥࢆ▱ࡿࡇࡀ㠀ᖖ㔜せ࡛࠶ࡿࠋ⌧Ⅼ࡛ࡣከࡃ ࡢୗ᮲㫅㢮ࡸཎጞⓗ࡞┿㦵㨶㢮㛵ࡋ࡚ࠊࢤࣀ ࣒ሗࡀ᫂ࡽ࡞ࡗ࡚࠾ࡽࡎࠊࡉࡽࡣྂࡃศ ᒱࡋࡓ㨶㢮࡛ࡣ࣮࢜ࢯࣟࢢ㑇ఏᏊ࡛ࡶ✀㛫࡛㓄ิࡀ ࠾࠾ࡁࡃ␗࡞ࡗ࡚࠸ࡿࡓࡵ PCR ࡼࡗ࡚ ancV1R ࡞ ࡢႥぬཷᐜయ㑇ఏᏊ⩌ࡢ༢㞳ࡍࡿࡇࡣᴟࡵ࡚ᅔ 㞴࡛࠶ࡿࠋࡑࡇ࡛ࠊᮏᨭ࠾࠸࡚ࠊࡇࢀࡲ࡛ࢤ ࣀ࣒㓄ิࡢሗ࿌ࡀ࡞࠸ࢢ࣮ࣝࣉࡘ࠸࡚ࠊႥୖ⓶ࡢ ࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒࢆᨭࡋ࡚࠸ࡓࡔࡁࡓ࠸ࠋ ᑐ㇟✀ࡋ࡚ࡣࠊୗ᮲㫅㢮ࡽࢳࣙ࢘ࢨ࣓ࠊࢫ࣏ ࢵࢸࢵࢺ࣮࢞ࠊࡑࡋ࡚ཎጞⓗ࡞┿㦵㨶㢮ࡽࠊࣟ ࣡ࢼࠊࣃࣥࢺࢻࣥࠊ࣑ࣔࣝࣝࢫࠊࢼࣇࣇࢵࢩࣗ ࢆィ⏬ࡋ࡚࠸ࡿࠋ⌧ẁ㝵࡛ࡣࠊྛ 1 ✀ࡘ࠸࡚ 1 ಶ యࡢႥୖ⓶⏤᮶ total RNA ࡢ㓄ิሗࢆ Hiseq2500 ࡼࡾ 1 ࣮ࣞࣥศ㸦PE㸧ྲྀᚓࡋ࡚࠸ࡓࡔࡁࡓ࠸⪃࠼ ࡚࠸ࡿࠋࡲࡓࠊཧ↷㓄ิࡀ࡞࠸ࡇࡽ TRINITY ࡼࡿ de novo ࢵࢭࣥࣈࣝࢆᐇࡋ࡚࠸ࡓࡔࡃணᐃ ࡛࠶ࡿ㸦ᮾᕤ࣭ఀ⸨ GP ౫㢗㸧 ࠋ ⾲䠍 ⬨᳝ື≀䝀䝜䝮୰䛻䛚䛡䜛㼍㼚㼏㼂㻝㻾㑇ఏᏊ䛾᭷↓ ့ ங 㝣 㢮 ᳇ ⏕ ≀ Ỉ ྂ ᳇ ௦ ⏕ 㨶 ≀ 㢮ே⊷䞉䝠䝖 ᪧୡ⏺䝄䝹 ᪂ୡ⏺䝄䝹 䛢䛳ṑ㢮 അ㋟㢮 㪒㢮 ወ㋟㢮 㣗⫗㢮 ⩼ᡭ㢮 ㈋ṑ㢮 㛗㰯㢮 䝝䜲䝷䝑䜽䝇㢮 ⿕⏥㢮䠄䜰䝹䝬䝆䝻䠅 ᭷⿄㢮 ༢Ꮝ㢮 㽢 㽢 䕿 䕿 䕿 㽢 䕿 䕿 㽢 䕿 䕿 䕿 䕿 䕿 䕿 㫽㢮 ∐㢮䠄䝖䜹䝀䠅 ୧⏕㢮 㽢 䕿 䕿 䝅䞊䝷䜹䞁䝇䠄⫗㫅㢮䠅 䕿 䝇䝫䝑䝔䝖䜺䞊 䕿 䝫䝸䝥䝔䝹䝇 䕿 ┿㦵㨶㢮 㽢 ᅗ㸯.⬨᳝ື≀ࡢ V1R 㑇ఏᏊࡢ⣔⤫ᶞ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ⌧ẁ㝵࠾࠸࡚ୖグࡢ㨶✀ࡘ࠸࡚Ⴅୖ⓶ࡽ total RNA ࢆᢳฟࡋ࡚࠸ࡿࡇࢁ࡛࠶ࡾࠊࡇࢀࡽᨭࢆ ᐇࡋ࡚࠸ࡓࡔࡃணᐃ࡛࠶ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᮏᨭࡼࡗ࡚ୗ᮲㫅㢮࠾ࡼࡧཎጞⓗ࡞┿㦵㢮 ࠾ࡅࡿ ancV1R ࡢ᭷↓ࡀ᫂ࡽ࡞ࡿඹࠊࡇࢀࡽ ࡢ㨶✀࠾࠸࡚ࡣࡢࡼ࠺࡞Ⴅぬཷᐜయ㑇ఏᏊࡀᏑ ᅾࡋࠊࡑࢀࡽࡀⓎ⌧ࡋ࡚࠸ࡿࡢࢆ᫂ࡽࡍࡿࡇ ࡀ࡛ࡁࡿࠋࡑࢀ௨እࡶࠊୗ᮲㫅㢮ࡽ┿㦵㢮 㐍ࢆࡋ࡚࠸ࡃ࠶ࡓࡾࠊႥぬཷᐜయ㑇ఏᏊ㸦OR, V1R, V2R, TAAR ࡞㸧ࡢࣞࣃ࣮ࢺ࣮ࣜࡀࡢࡼ࠺ ኚࡋ࡚࠸ࡗࡓࡘ࠸࡚ࡶࠊ᫂ࡽࡍࡿࡇࡀ ࡛ࡁࡿᮇᚅࡋ࡚࠸ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ⌧ᅾࡢࡇࢁࠊࡲࡔᨭෆᐜࢆㄽᩥࡋ࡚බ⾲ࡋ࡚ ࠸࡞࠸ - 463 - බເᨭㄢ㢟㸸2013 ᖺᗘ ୗ⑩ᝈඣ⏤᮶ࡢ⭠⳦࠾ࡅࡿཎ㑇ఏᏊ࣭⸆⪏ᛶ㑇ఏᏊࡢࣃࣥࢤࣀ࣒ゎᯒ ᇶ┙◊✲㸦㹁㸧 す 㡰୍㑻࣭㮵ඣᓥᏛᏛ㝔་ṑᏛ⥲ྜ◊✲⛉࣭ᚤ⏕≀Ꮫศ㔝 㸺⫼ᬒ࣭┠ⓗ㸼 ⭠ ⟶ จ 㞟 ᛶ ⭠ ⳦ 㸦 Enteroaggregative Escherichia coli, EAEC㸧ࡣ HEp-2 ⣽⬊╔ヨ㦂࡛ จ㞟ᛶ╔ࢆ♧ࡍ⭠⳦ᐃ⩏ࡉࢀࠊ⭠⟶⢓⭷ biofilm ࢆᙧᡂࡋࠊᑠඣ࣭ᡂேࡢୗ⑩ࡢཎᅉ࡞ࡿࠋ ୍⯡ࡢ⮫ᗋ᳨ᰝᐊ࡛ࡣ᳨ᰝࡉࢀ࡚࠾ࡽࡎࠊ⮫ᗋⓗ㔜 せᛶࡀ༑ศホ౯ࡉࢀ࡚࠸࡞࠸ࠋEAEC ࡢ PCR ࡼࡿ ྠᐃࡣ㌿ᅉᏊ AggR ࡢ㑇ఏᏊ㸦aggR㸧ࡀ⏝࠸ࡽ ࢀ࡚࠸ࡿࡀࠊaggR ࢆᣢࡓ࡞࠸ atypical EAEC ࡶከ ᩘࡳࡽࢀࡿࠋEAEC ࡣ㑇ఏⓗከᵝᛶࡀࡳࡽࢀࡿࡀࠊ ከᩘࡢᰴࢆᑐ㇟ࡋࡓ⣔⤫ゎᯒࡢሗ࿌ࡣᑡ࡞࠸ࠋࡲ ࡓࠊEAEC ࡢࢤࣀ࣒ゎㄞࡣࣉࣟࢺࢱࣉ 042 ᰴ࡞ 2 ᰴࡔࡅ࡛࠶ࡾࠊከࡃࡢ EAEC ᰴࢆᑐ㇟ࡋࡓࢤࣀ ࣒ẚ㍑ࡣ⾜ࢃࢀ࡚࠸࡞࠸ࠋᮏ◊✲࡛ࡣ⣔⤫ゎᯒࢤ ࣀ࣒ゎᯒࢆ㏻ࡌ࡚ࠊࢃࡀᅜࡢ EAEC ࡢከᵝᛶ≉ᚩ ࢆ᫂ࡽࡍࡿࡇࢆ┠ⓗࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ᑐ㇟ࡣ㮵ඣᓥ┴ࡢᑠඣୗ⑩ᝈඣ⏤᮶ࡢ aggR 㝧 ᛶ⭠⳦ 138 ᰴ atypical EAEC 22 ᰴ࠾ࡼࡧ㜰 ᗓබ⾗⾨⏕◊✲ᡤ࡛ศ㞳ࡉࢀࡓ aggR 㝧ᛶ⭠⳦ 31 ᰴࠋAtypical EAEC ࡣ biofilm assay ࡛ࢫࢡ࣮ࣜࢽ ࣥࢢࡋࠊHEp-2 ⣽⬊╔ヨ㦂࡛☜ᐃࡋࡓᰴࢆ⏝࠸ࡓࠋ ᑐ㇟ᰴࡢཎ㑇ఏᏊࠊᇶ㉁ᣑᙇᆺșࣛࢡࢱ࣐࣮ࢮ 㸦ESBL㸧㑇ఏᏊ blaCTX-M ࠊ⾑ΎᆺࢆㄪࠊMLSA (multilocus sequence analysis㸧ࢆ⾜ࡗࡓࠋࡉࡽࠊ ࣑ࣝࢼ MiSeq ࡛⣔⤫ࡢ␗࡞ࡿ 25 ᰴࢻࣛࣇࢺ㓄ิ ࢆỴᐃࡋࠊせ࡞ཎᅉᏊࡢศᕸࢆ blast ᳨⣴ࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 MLSA ࡼࡿ⣔⤫ゎᯒ࡛ࡣࠊaggR 㝧ᛶ⭠⳦ࡣ ⭠⳦ࡢ⣔⤫⩌ AࠊB1ࠊB2ࠊD ࡢ 4 ࡘᗈࡃศᕸࡋ࡚ ࠸ ࡓ 㸦 ᅗ 1 㸧ࠋ ST200 (O126:H7 ࡞ ) ࠊ ST40 (O111:H21 ࡞)ࠊST3570 (O86a:H27 ࡞)ࡣࠊ㮵 ࣉࣛࢫ࣑ࢻࡢ⬺ⴠࡢྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ ඣᓥ㜰ࡢ୧᪉࡛ศ㞳ࡉࢀࠊࡲࡓ 1990 ᖺ௦ᚋ༙ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡣࡍ᳨࡛ฟࡉࢀ࡚࠾ࡾࠊࢃࡀᅜ௨๓ࡽࡳࡽࢀ ࢃࡀᅜ࡛ศ㞳ࡉࢀࡿ EAEC ࡢ⣔⤫ゎᯒࢆึࡵ࡚ᐇ ࡿせ࡞ EAEC ࡛࠶ࡿ᥎ᐃࡉࢀࡿࠋ ࡛ࡁࠊࢃࡀᅜ࡛ᗈࡀࡗ࡚࠸ࡿせ࡞ࢡ࣮ࣟࣥࡢᏑ ୍᪉ࠊaggR 㝧ᛶ࡛ B2 ᒓࡍࡿ O25:H4:ST131 ࡢ ᅾࢆ☜ㄆ࡛ࡁࡓࠋࡲࡓࠊ⤒ᖺⓗࡳࡿࡇ࡛ࠊ10 ᩘ ࢡ࣮ࣟࣥࡣࠊ2003 ᖺ௨ᚋ㮵ඣᓥ࡛ฟ⌧ࡋ࡚࠾ࡾ㸦ᅗ ᖺࡢ⠊ᅖ࡛ࢡ࣮ࣟࣥࡢኚ㑄ࢆ᫂ࡽࡍࡿࡇࡀ࡛ 1㸧ࠊࡇࢀࡽࡢᰴࡣྂⓗ࡞ EAEC ࡀಖ᭷ࡍࡿᰁⰍయ ࡁࡓࠋࡃ UPEC ࡀ EAEC ࡢཎ㑇ఏᏊ ESBL ⏤᮶ࡢ EAEC ཎ㑇ఏᏊ picࠊshET1ࠊaaiC ࢆᣢࡗ (CTX-M)㑇ఏᏊࢆ⋓ᚓࡋ࡚ᗈࡀࡗ࡚࠸ࡿࡇࡀࢃ ࡚࠾ࡽࡎࠊᒀ㊰ཎᛶ⭠⳦(UPEC)ࡢ╔㑇ఏᏊ ࡾࠊ⭠⟶እཎᛶ⭠⳦ୗ⑩ཎᛶ⭠⳦ࡢ㛫࡛ࠊ afaB ࢆᣢࡗ࡚࠸ࡓ㸦ᅗ 2㸧ࠋࡋࡓࡀࡗ࡚ࠊୡ⏺ⓗ ཎ㑇ఏᏊ࣭⸆⪏ᛶ㑇ఏᏊࡢỈᖹఏࡀⴭ᫂㐍 ᘏࡋ࡚࠸ࡿ UPEC O25:H4:ST131 ࡀ EAEC ࡢཎ ࢇ࡛࠸ࡿࡇࡀ♧၀ࡉࢀࡓࠋจ㞟ᛶࡢᙉ࠸ EAEC ࡀ ࣉࣛࢫ࣑ࢻࢆ⋓ᚓࡋ࡚ฟ⌧ࡋࡓࡇࡀ♧၀ࡉࢀࡓࠋ ࡇࢀࡽࡢ㑇ఏᏊࡢỈᖹఏࡢࡼ࠺㛵ࢃࡗ࡚࠸ ࡉࡽࡇࡢࢡ࣮ࣟࣥࡣ blaCTX-M-14 ࢆಖ᭷ࡋ࡚࠾ࡾࠊ ࡿࡢࠊᚋࡢ᪂ࡓ࡞◊✲ㄢ㢟࡛࠶ࡿࠋࡲࡓࠊEAEC ESBL ࢆ⏘⏕ࡋ࡚࠸ࡿࡇࡶ᫂ࡽ࡞ࡗࡓࠋ ࡢྠᐃࡣ aggR ࡔࡅ࡛ࡣ༑ศ࡛࠶ࡾࠊEAEC ࡢ Atypical EAEC ࡣ⣔⤫⩌ A B2 ศᕸࡋࠊO ⾑ ᐃ⩏ᇶ࡙࠸ࡓ᳨ฟἲࡢ᳨ウࡶᚲせ࡞ࡗࡓࠋ Ύ⩌ࡣ UT (untypable)ࡀከࡗࡓࠋࢤࣀ࣒ゎᯒ࡛ࡣ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 EAEC ≉᭷࡞ཎ㑇ఏᏊࡣࡳࡽࢀࡎࠊྂⓗ࡞ EAEC ࡣࡃ␗࡞ࡿ⣔⤫࡛࠶ࡿࡇࡀࢃࡗࡓࠋ Phylogenetic analysis of EAEC isolated in Kagoshima and Osaka, Japan, revealed aggR 㝧ᛶᰴࡋ࡚ࢤࣀ࣒ゎᯒࢆᐇࡋࡓᰴࡢ୰ emergence of CTX-M-14-producing EAEC ࡣࠊaggR ࠾ࡼࡧ EAEC ࡢࡑࡢࡢཎ㑇ఏᏊࡀ O25:H4:ST131. Imuta N. et al (in submission) ࢇࡳࡽࢀ࡞࠸ᰴࡶ࠶ࡾࠊPCR ࡢ㝧ᛶࡸཎ - 464 - බເᨭㄢ㢟㸸2014 ᖺᗘ ⳦㢮ࡢ࢚࢟ࢯࣥࠊࣥࢺࣟࣥᵓ㐀ࡢ㐍ᶵᵓࡢゎ᫂ ᇶ┙◊✲㸦㹁㸧 す⏣ ὒᕭ࣭ᐩᒣ┴❧ᏛᕤᏛ㒊 㸺⫼ᬒ࣭┠ⓗ㸼 ⳦㢮ࡢ࢚࢟ࢯ࣭ࣥࣥࢺࣟࣥᵓ㐀ࡣከᵝᛶࡀぢ ࡽࢀࡿࠋࡍ࡞ࢃࡕࠊᢸᏊ⳦㢮㸦Basidiomycota㸧࡛ࡣࠊ ࣥࢺࣟࣥᩘࡀࡢ⣔⤫⩌ࡼࡾࡶከࡃࠊࣥࢺࣟࣥ ࢧࢬࡣᑠࡉ࠸ࠋ᪉ࠊᏊᄞ⳦㓝ẕ㸦Saccharomycotina㸧 ࡛ࡣࠊࣥࢺࣟࣥᩘࡀᑡ࡞ࡃࠊࣥࢺࣟࣥࢧࢬࡣ ✀ࡼࡗ࡚ᖜࡀ࠶ࡿࠋ⣒≧Ꮚᄞ⳦㢮㸦Pezizomycotina㸧 ࡣᢸᏊ⳦㢮Ꮚᄞ⳦㓝ẕࡢ୰㛫ⓗ࡞≉ᚩࢆᣢࡗ࡚࠸ ࡿࠋࡋࡋࠊᏊᄞ⳦㢮㸦Ascomycota㸧ࢆᵓᡂࡋ࡚࠸ ࡿ ⣔ ⤫ ⩌ ࡢ ୍ ࡘ ࡛ ࠶ ࡿ ྂ ⏕ Ꮚ ᄞ ⳦ 㢮 㸦 Taphrinomycotina 㸧 ࡢ ࢤ ࣀ ࣒ ሗ ࡣ ศ 㓝 ẕ Schizosaccharomyces ᒓ⳦✀㝈ࡽࢀ࡚࠾ࡾࠊࡇࡢ⣔ ⤫⩌ࡢࢤࣀ࣒ࡢከᵝᛶࡸ≉ᚩࡘ࠸࡚ࡣ࡛᫂࠶ࡿࠋ ࡇࢀࡲ࡛ࡢ◊✲ࡼࡾࠊ࢚࢟ࢯࣥ㡿ᇦࡣࣥࢺࣟ ࣥ㡿ᇦࡼࡾࡶሷᇶ⤌ᡂ࠾ࡅࡿࢢࢽ࣭ࣥࢩࢺࢩࣥ 㸦GC㸧ྵ㔞ࡀ㧗ࡃࠊࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂ㢖ᗘࡶ㧗࠸ ࡇࡀ♧ࡉࢀ࡚࠸ࡿࠋࡑࡇ࡛ࠊᮏ◊✲࡛ࡣࠊྂ⏕Ꮚ ᄞ⳦㢮ᒓࡍࡿ Saitoella complicata ࡢࢤࣀ࣒ DNAࠊ Ⓨ⌧ RNAࠊࢾࢡࣞ࢜ࢯ࣮࣒ DNA ᩿∦ࢆࡑࢀࡒࢀࢩ ࣮ࢡ࢚ࣥࢫࡋࠊS. complicata ࠾ࡅࡿࢤࣀ࣒ࡢ≉ᚩ Schizosaccharomyces pombe ࡢ≉ᚩࢆẚ㍑ࡍࡿࡇࢆ ┠ⓗࡋࡓࠋ࡞࠾ࠊS. complicata ࡣศ࡛ࡣ࡞ࡃࠊฟ ⱆ࡛ቑṪࡍࡿࡀࠊ⣽⬊⾲ᒙࡣᏊᄞ⳦㓝ẕᆺࢆࡋ࡚࠸ ࡿࡶࢃࡽࡎࠊෆ⏕ศⱆᆺ㸦enteroblastic㸧ࡢᢸ Ꮚ⳦㓝ẕ࡛ぢࡽࢀࡿฟⱆᵝᘧࢆᣢࡗ࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 S. complicata ࡢࢤࣀ࣒ DNA ࡢࢩ࣮ࢡ࢚ࣥࢫࡢᨭ 㸦ᯘဴஓඛ⏕࣭ᑠ᳚⩏ಇඛ⏕㸧ࠊࣄࢫࢺࣥ⬺ࢭࢳ ࣝ㜼ᐖࢺࣜࢥࢫࢱࢳࣥ A㸦TSA㸧ࢆ 1 ȝJP/ ȝJP/ ȝJP/ ຍ࠼ࡓ᮲௳࡛ᇵ㣴ࡋࡓ S. complicata ࡼࡾᢳฟࡋࡓ RNA ࡢࢩ࣮ࢡ࢚ࣥࢫࡢᨭ㸦㕥ᮌ✨ ඛ⏕㸧ࠊTSA ࢆ 0 ȝJP/㸦ࢥࣥࢺ࣮ࣟࣝ㸧ȝJP/ ຍ ࠼ࡓ᮲௳࡛ᇵ㣴ࡋࡓ S. complicata ࡼࡾᢳฟࡋࡓ༢୍ ࢾࢡࣞ࢜ࢯ࣮࣒ DNA ᩿∦ࡢࢩ࣮ࢡ࢚ࣥࢫࡢᨭ 㸦㕥ᮌ✨ඛ⏕㸧ࢆཷࡅࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 S. complicata ࡢࢤࣀ࣒ DNA ࢩ࣮ࢡ࢚ࣥࢫࡼࡾࠊ 104 ࢥࣥࢸࢢ㸦N50, 239 Kbp㸧ࠊ㛗 14.1 Mbp ࢆỴ ᐃࠊබ㛤ࡋࡓࠋࡇࢀࡣྂ⏕Ꮚᄞ⳦㢮࠾࠸࡚ࠊ Schizosaccharomyces ᒓ⳦✀௨እ࡛ึࡵ࡚ࡢࢤࣀ࣒ ሗ࡞ࡿࠋࡲࡓࠊRNA ࣐ࢵࣉሗࢆ⪃៖ࡋࠊ6933 ࡢ ࢱࣥࣃࢡ㉁ࢥ࣮ࢻ㡿ᇦࠊ7119 ࡢ㌿ࣘࢽࢵࢺࢆ᫂ࡽ ࡋࠊබ㛤ࡋࡓࠋࢤࣀ࣒ሷᇶ㓄ิ࠾ࡼࡧ㑇ఏᏊὀ 㔘ࡅࡼࡗ࡚ࠊ࢚࢟ࢯࣥ㡿ᇦ㸦n = 20524㸧ࡢᖹᆒ GC ྵ㔞ࡀ 52.4%ࠊࣥࢺࣟࣥ㡿ᇦ㸦n = 13591㸧ࡢᖹ ᆒ GC ྵ㔞ࡀ 48.0%࡛࠶ࡿࡇࡀࢃࡗࡓࠋ ࢾࢡࣞ࢜ࢯ࣮࣒ࢆᙧᡂࡋ࡚࠸ࡿ DNA ᩿∦࠾ࡅ ࡿ 2 ሷᇶ㓄ิࣃࢱ࣮ࣥ㸦ࢤࣀ࣒ࢩࢢࢿࢳࣕ㸧ࢆẚ㍑ ࡋࡓࡇࢁࠊS. complicata ࡣ Sch. pombe ࡁࡃ㐪ࡗ ࡚࠸ࡿࡇࢆ᫂ࡽࡋࡓࠋS. complicata ࡢࡑࢀࡣࠊ ᢸᏊ⳦㢮࡛࠶ࡿ Mixia osmundae ࡢ 2 ሷᇶ㓄ิࣃࢱ࣮ ࣥࡢ᪉㢮ఝࡋ࡚࠸ࡓࠋࡇࡢ⤖ᯝࡣࠊࢾࢡࣞ࢜ࢯ࣮ ࣒ᙧᡂࡣྛ┿᰾⏕≀✀࡛㧗ᗘ⥔ᣢࡉࢀ࡚࠾ࡾࠊ㐍 ⣔⤫ୖ㏆⦕࡞⏕≀✀㛫࡛࠶ࡗ࡚ࡶࡑࡢไᚚࡀ␗࡞ ࡗ࡚࠸ࡿࡇࢆ♧ࡋ࡚࠸ࡿࠋS. complicata ࡢࢤࣀ࣒ GC ྵ㔞ࡀ 52.6%࡛࠶ࡾࠊSch. pombe ࡢ 36.0%ࡁ ࡃ㐪ࡗ࡚࠾ࡾࠊM. osmundae ࡢ 55.5%㏆࠸ࡇ㛵 㐃ࡀ࠶ࡿ⪃࠼ࡽࢀࠊ⏕≀⣔⤫ࡼࡾࡶࢤࣀ࣒ሷᇶ⤌ ᡂࡢ᪉ࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂࡀᙉࡃ㛵㐃ࡋ࡚࠸ࡿࡇ ࢆ♧၀ࡋ࡚࠸ࡿࠋ TSA ⃰ᗘࡀ 3 ȝJP/ ࡢሙྜࡣࠊ᫂ࡽ࡞ฟⱆ␗ ᖖࡀ㢧ᚤ㙾ࡼࡾほᐹࡉࢀࡓࡶࢃࡽࡎࠊTSA ⃰ᗘ౫Ꮡⓗ㑇ఏᏊⓎ⌧ࡀ 25%௨ୖቑຍࡋࡓ㑇ఏᏊ ᩘࡣ 154ࠊῶᑡࡋࡓ㑇ఏᏊᩘࡣ 131 ᴟࡵ࡚ᑡ࡞ࡗ ࡓࠋࡇࢀࡽ㑇ఏᏊⓎ⌧ኚࡀ⏕ࡌࡓ㑇ఏᏊ࠾ࡅ ࡿࢾࢡࣞ࢜ࢯ࣮࣒⨨ࡢኚࡶࢇぢࡿࡇࡀ ࡛ࡁ࡞ࡗࡓࠋࡲࡓࠊTSA ⃰ᗘࡀ 3 ȝJP/ ࡢࡁࡢ ࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂ㡿ᇦࡢ 2 ሷᇶ㓄ิࣃࢱ࣮ࣥࢥ ࣥࢺ࣮ࣟࣝࢆẚ㍑ࡋ࡚ࡶ㢧ⴭ࡞㐪࠸ࡣぢࡽࢀ࡞ࡗ ࡓࠋࡇࢀࡽࡢࡇࡣࠊࣄࢫࢺࣥࢭࢳࣝࡢஹ㐍 ࡼࡿࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂኚࡣᴟࡵ࡚㝈ࡽࢀ࡚࠾ࡾࠊ S. complicata ࠾ࡅࡿࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂࣃࢱ࣮ࣥ ࡣ㧗ᗘಖᏑࠊ⥔ᣢࡉࢀ࡚࠸ࡿࡇࢆ♧ࡋ࡚࠸ࡿࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 S. complicata ࡢࢤࣀ࣒ሷᇶ㓄ิ࠾ࡼࡧ㑇ఏᏊὀ㔘 ࡣබ㛤ࠊⓎ⾲ࡋࡓࡀࠊRNA ࣐ࢵࣉ࠾ࡼࡧࢾࢡࣞ࢜ࢯ ࣮࣒࣐ࢵࣉゎᯒࡢ⤖ᯝࡣ⌧ᅾㄽᩥᢞ✏୰࡛࠶ࡿࠋ ┿᰾⏕≀࠾࠸࡚ࠊࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂࡣ⣽⬊ࡢ ⥔ᣢࡢࡓࡵᴟࡵ࡚㔜せ࡛࠶ࡿࠋࡋࡋࠊᅇࡢᐇ 㦂⤖ᯝࡼࡗ࡚ࠊ㐍⣔⤫ࡀ㏆࠸ࡶࢃࡽࡎࠊ ࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂᵝᘧ㸦ࣄࢫࢺࣥ DNA ሷᇶ㓄ิ ࡢ㛵ಀ㸧ࡀ㐪࠸ࠊ⣔⤫ⓗ㞳ࢀ࡚࠸࡚ࡶࢤࣀ࣒ሷ ᇶ⤌ᡂࡀఝ࡚࠸ࡿሙྜࠊࢾࢡࣞ࢜ࢯ࣮࣒ᙧᡂᵝᘧ ࡀఝ࡚࠸ࡿࡇࡀࢃࡗࡓࠋࡇࢀࡣࠊ㐍࠾࠸࡚ ✚ࡋ࡚࠸ࡿ DNA ࡢሷᇶ⨨㸦✺↛ኚ␗㸧ࢾࢡࣞ ࢜ࢯ࣮࣒ᙧᡂࡀ㛵㐃ࡋ࡚࠾ࡾࠊሷᇶ⤌ᡂ౫Ꮡⓗࣄ ࢫࢺࣥ DNA ࡢ┦స⏝ࡀ⏕ࡌ࡚࠸ࡿࡇࢆព ࡋ࡚࠸ࡿࠋ┿᰾⏕≀࠾ࡅࡿ DNA ࡢሷᇶ㓄ิࡢᶵ ⬟ⓗ࡞ពࢆゎ᫂ࡍࡿ⣒ཱྀ࡞ࡿ☜ಙࡍࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Yamauchi K et al. (2015) Genome Announcements 3:e00220-15 - 465 - බເᨭㄢ㢟㸸2010 ᖺᗘ ⫾ඣᆺ⭈⮚ᖿ⣽⬊ࡢᡂయ⭈࡛ࡢάᛶ ᣮᡓⓗⴌⱆ◊✲ す୰ᮧ 㝯୍࣭⇃ᮏᏛⓎ⏕་Ꮫ◊✲ᡤ 㸺⫼ᬒ࣭┠ⓗ㸼 ⣒⌫యࡸᒀ⣽⟶ࡽ࡞ࡿ⭈⮚ࡢ᭱ᑠᶵ⬟༢ࢆࢿ ࣇࣟࣥࡪࠋᡂయࡢ⭈⮚ࡣ⏕ࡋ࡞࠸⮚ჾ࡛࠶ࡿ ࡀࠊ⫾ඣᮇࡣࢿࣇࣟࣥࡀ๓㥑⣽⬊ࡽᙧᡂࡉࢀࡿࠋ ࡋࡋࡇࡢࢿࣇࣟࣥ๓㥑⣽⬊ࡣࠊ࣐࢘ࢫ࡛ࡣ⏕┤ᚋ ࠊࣄࢺ࡛ࡣㄌ⏕๓ࠊࡍ࡚ศࡋ࡚ᾘኻࡋ࡚ࡋ ࡲ࠺ࠋᡂయ⭈⮚ෆ࡛ࢿࣇࣟࣥ๓㥑⣽⬊ࢆάᛶ࡛ ࡁࢀࡤࠊ᪂ࡓ࡞ࢿࣇࣟࣥࡢᙧᡂࡀྍ⬟࡞ࡿࡶࡋ ࢀ࡞࠸ࠋࡑࡢࡓࡵࡣࢿࣇࣟࣥ๓㥑⣽⬊࡛ാࡃ㌿ ᅉᏊࢿࢵࢺ࣮࣡ࢡࡑࡢᶆⓗྠᐃࡀᚲ㡲࡛࠶ࡿࠋ ᡃࠎࡣࠊ ᰾ෆᅉᏊ Sall1 ࡀ⭈⮚Ⓨ⏕ᚲ㡲࡞ࡇࠊ ࢿࣇࣟࣥࡀࡑࡢඖ࡞ࡿ⣽⬊⩌㸦ࢿࣇࣟࣥ๓㥑⣽⬊㸧 ࡽ⏤᮶ࡋࠊࡇࡢࢿࣇࣟࣥ๓㥑⣽⬊ཬࡧศึᮇࡢ ࢿࣇࣟࣥ Sall1 ࡀⓎ⌧ࡍࡿࡇࢆሗ࿌ࡋ࡚ࡁࡓ (Nishinakamura et al., Development 2001; Osafune et al., Development 2006)ࠋࢿࣇࣟࣥ๓㥑 ⣽⬊ࡀᮍศࡢࡲࡲ⮬ᕫ」〇ࡋ࡞ࡀࡽᑡࡋࡎࡘࢿࣇ ࣟࣥศࡋ࡚࠸ࡃࡇࡀࠊ⭈⮚ᙧᡂᴟࡵ࡚㔜せ ࡛࠶ࡿࠋࡇࡢࣂࣛࣥࢫࡢไᚚࡣࠊWnt ࢩࢢࢼࣝ ࡼࡿศಁ㐍㌿ᅉᏊ Six2 ࡼࡿᢚไࡀ㛵ࡋ࡚ ࠸ࡿࡇࡀࢢ࣮ࣝࣉࡽሗ࿌ࡉࢀ࡚࠸ࡿࡀࠊ౫↛ ᮍゎ᫂࡞Ⅼࡀከࡃࠊࢿࣇࣟࣥ๓㥑⣽⬊࠾ࡅࡿ Sall1 ࡢᙺࡶ㛗࠸㛫ㅦࡢࡲࡲࡔࡗࡓࠋࡑࡇ࡛ᮏ◊✲࡛ࡣࠊ ᡂయ⭈࡛ࡢάᛶྥࡅ࡚ࡢᇶ┙ࡋ࡚ࠊ⫾ඣᮇ ࢿࣇࣟࣥ๓㥑⣽⬊࠾ࡅࡿ Sall1 Six2 ࡢᶆⓗ㑇ఏ Ꮚ⩌ࢆྠᐃࡋࠊࡑࡢไᚚᶵᵓࢆゎ᫂ࡍࡿࡇࢆ┠ⓗ ࡋࡓࠋ 㸺ᨭࡢෆᐜ㸼 ࢿࣇࣟࣥ๓㥑⣽⬊ࡢ⥔ᣢศࢆไᚚࡍࡿSall1 Six2ࡢศᏊᶵᵓࢆゎ᫂ࡍࡿࡓࡵࠊⓎ⏕ᮇࡢ⭈⮚ࢆ ⏝࠸࡚Sall1ᢠయཬࡧSix2ᢠయࡼࡿࢡ࣐ࣟࢳࣥච ỿ㝆ࢩ࣮ࢡ࢚ࣥࢫ (ChIP-seq) ࢆ⾜ࡗࡓࠋලయⓗ ࡣᡃࠎࡀ࣐࢘ࢫ⫾⏕16.5᪥ࡢ⭈⮚ࢆ⏝࠸࡚ChIP ࢆ⾜࠸ࠊỿ㝆DNAࢆᮾிᏛࡢⳢ㔝◊✲ᐊ㏦ࡗ࡚ࠊ 㧗㏿ࢩ࣮ࢡ࢚ࣥࢧ࣮ࡼࡿࢩ࣮ࢡ࢚ࣥࢫゎᯒཬࡧ࣐ ࢵࣆࣥࢢࢆᨭࡋ࡚࠸ࡓࡔ࠸ࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᡃࠎࡣࡲࡎSall1ࡀࢿࣇࣟࣥ๓㥑⣽⬊ࡢ⮬ᕫ」〇 ⥔ᣢᚲ㡲࡛࠶ࡿࡇࢆぢ࠸ࡔࡋࡓࠋࢿࣇࣟࣥ๓㥑 ⣽⬊≉␗ⓗSall1ࢆࣀࢵࢡ࢘ࢺࡋࡓࡇࢁࠊ⭈⮚ ࡣᑠࡉࡃ࡞ࡗ࡚ࢿࣇࣟࣥࡶࡰᾘኻࡋࡓࠋࢿࣇࣟࣥ ๓㥑⣽⬊ࡢ⮬ᕫ」〇⬟ࡀపୗࡋ࡚ศࡋ࡚ࡋࡲ࠸ࠊ ࡉࡽࡑࡢศࡋࡓึᮇࢿࣇࣟࣥࡶ⣽⬊Ṛࢆ㉳ࡇࡋ ࡚ࡋࡲ࠺ࡓࡵ࡛࠶ࡗࡓࠋḟ࠸࡛ࠊ⸆ㄏᑟᛶࡢSall1 ࣀࢵࢡ࢘ࢺ࣐࢘ࢫࢆస〇ࡋࠊ࣐ࢡࣟࣞゎᯒ ࡼࡗ࡚ࠊSall1ࡀࠊࢿࣇࣟࣥ๓㥑⣽⬊࠾࠸࡚ࡣୗ ὶ㑇ఏᏊࡢ㌿ࢆಁ㐍ࡋ࡚ᮍศ≧ែࢆ⥔ᣢࡋࠊศ ୰ࡢึᮇࢿࣇ࡛ࣟࣥࡣ ␗ᖖ࡞㌿ࢆᢚไࡍࡿࡇ ࡀุ᫂ࡋࡓࠋࡉࡽChIP-seqゎᯒࡼࡗ࡚ࠊSall1 Six2ࡢᶆⓗ㑇ఏᏊࡀ࡞ࡾ㔜」ࡋࠊࡘ⤖ྜ㒊 ࡀ㏆ഐ (500bp௨ෆ) ⨨ࡍࡿࡶࡢࡀከ࠸ࡇࡀ ᫂ࡽ࡞ࡗࡓࠋࡘࡲࡾࢿࣇࣟࣥ๓㥑⣽⬊࠾࠸࡚ Sall1ࡣSix2ࡶࠊ㔜せ࡞⭈⮚ᙧᡂ㑇ఏᏊ⩌ (Osr1, Robo2, Eya1࡞) ࡢไᚚ㡿ᇦ┤᥋⤖ྜࡋ ࡚࠸ࡓࠋSall1ࡢ⤖ྜࡍࡿࢥࣥࢭࣥࢧࢫ㓄ิࡶྠᐃࡉ ࢀࡓࠋࡲࡓSall1Six2ࡣࢱࣥࣃࢡྠኈ࡛ࡶ⤖ྜࡋࡓࠋ ୍᪉ࠊSix2ࡀᏑᅾࡋ࡞࠸ึᮇࢿࣇࣟࣥ࠾࠸࡚ࡣࠊ Sall1ࡣMi2/NuRD」ྜయ⤖ྜࡋ࡚㌿ࢆᢚไࡋ ࡚࠸ࡓࠋࡇࡢሙྜࡢᶆⓗ㑇ఏᏊ (Nkx6.1࡞) ࡣ Six2ࡢࡑࢀ (Wnt4,Fgf8࡞) ࡣ␗࡞ࡾࠊࡘไᚚ 㡿ᇦࡢ┤᥋⤖ྜࢆࡉ࡞࠸ࡇࡶุ᫂ࡋࡓࠋࡇࡢ ࡼ࠺Sall1ࡣࠊSix2୍㒊㔜」ࡍࡿࡶࡢࡢ␗࡞ࡿ࣓ ࢝ࢽࢬ࣒࡛ࠊࢿࣇࣟࣥ๓㥑⣽⬊࡛ࡣάᛶᅉᏊࠊึ ᮇࢿࣇ࡛ࣟࣥࡣᢚไᅉᏊࡋ࡚ࠊࡇࡢ㸰ࡘࡢᮍ⇍࡞ ⣽⬊㞟ᅋࢆ⥔ᣢࡋ࡚࠸ࡿࡇࡀゎ᫂ࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᅇࡢᡂᯝࡣࠊᡂయ⭈࠾࠸࡚ࢿࣇࣟࣥ๓㥑⣽⬊ ࢆάᛶࡍࡿ㝿ࡢᇶ┙ⓗ▱ぢ࡞ࡿࡇࡀᮇᚅࡉ ࢀࡿࠋࡲࡓᡃࠎࡣࣄࢺ iPS ⣽⬊ࡽࡢࢿࣇࣟࣥ๓㥑 ⣽⬊ࡢㄏᑟᡂຌࡋ࡚࠾ࡾ (Taguchi et al., Cell Stem Cell 2014)ࠊࣄࢺࢿࣇࣟࣥ๓㥑⣽⬊ࢆேⅭⓗ ቑᖜࡍࡿᢏ⾡㛤Ⓨࡗ࡚ࡶ᭷⏝࡛࠶ࡿࠋ ᇵ㣴⣽⬊ࢆࡗࡓ ChIP-seq ࡣ᪤࣮ࣝࢳ࣮ࣥ࣡ ࢡࡢ㡿ᇦ࡛࠶ࡿࠋࡋࡋⓎ⏕Ꮫࡣ㠀ᖖᚤ㔞ࡢࢧࣥ ࣉࣝࢆᢅ࠺㡿ᇦ࡛࠶ࡿࡓࡵࠊChIP-seq ࡢᡂᯝࡣᑡ࡞ ࠸ࠋᅇࡢᡂᯝࡣⓎ⏕Ꮫࢤࣀ࣒⛉Ꮫࡢ⼥ྜࡢ୍ ࡛࠶ࡾࠊᚋࠊ⮚ჾᙧᡂ࠾ࡅࡿ⥙⨶ⓗ࡞ศᏊࢿࢵ ࢺ࣮࣡ࢡゎ᫂ࡢ㐍ᒎࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Kanda, S. et al., J. Am. Soc. Nephrol. 25, 2584-2595 (2014) - 466 - බເᨭㄢ㢟㸸2015 ᖺᗘ ⾲⌧ᆺྍረᛶ㸸⏕ែᏛศᏊⓎ⏕Ꮫࡢ⤫ྜⓗ◊✲ ᇶ┙◊✲㸦㸿㸧 すᮧ Ḡஓ࣭ᾏ㐨ᏛᏛ㝔Ỉ⏘⛉Ꮫ◊✲㝔 㸺⫼ᬒ࣭┠ⓗ㸼 ⏕≀ࡀ⧊ࡾ࡞ࡍከᵝ࡞⾲⌧ᆺࡣ⏕≀✀ࡼࡿࢤࣀ࣒ ࡢከᵝᛶ౫ࡿゝࡗ࡚ࡶ㐣ゝ࡛ࡣ࡞࠸ࡀ㸪⎔ቃ ᛂࡌࡓ⾲⌧ᆺྍረᛶࡣ㸪ኚືࡍࡿ⎔ቃࡢ୰࡛⏕≀ࡀ ⏕ࡁᢤࡃࡓࡵ㠀ᖖ㔜せ࡞ࡶࡢ࡛࠶ࡿࡔࡅ࡛࡞ࡃ㸪 ✀ศࡢᘬࡁ㔠࡞ࡿྍ⬟ᛶࡀ࠶ࡿ㸬࢚ࢰࢧࣥࢩࣙ ࢘࢘࢜(Hynobius retardatus)ࡢᗂ⏕ࡣᤕ㣗࣭⿕㣗㸪 ➇த࡞ࡢ≧ἣ࡛⾲⌧ᆺྍረᛶࡼࡿᙧែ࣭⾜ືࡢ ከᆺࢆ♧ࡋ㸪ࡑࢀࡽࡢ⾲⌧ᆺࡣ⏕ែᏛⓗ㐺ᛂᶵ⬟ࢆ ᣢࡘࡓࡵ㸪⏕ែᏛࡢᑐ㇟ࡋ࡚◊✲ࡀ✚ࡉࢀ࡚ࡁ ࡓ㸬ࡋࡋ㸪ྍረᛶࡢⓎ⏕Ꮫⓗᇶ┙ࡘ࠸࡚ࡣ ࢇศࡗ࡚࠸࡞࠸㸬ᮏ⏦ㄳㄢ㢟࡛ࡣ㸪ᮏ✀ࢆᑐ㇟ ࡋ࡚⾲⌧ᆺྍረᛶ㛵ࡍࡿ㑇ఏᏊ㓄ิࣃࢱ࣮ࣥ ࡘ࠸࡚㸪ᆅᇦ㞟ᅋࡢ㞟ᅋ㑇ఏᏛⓗẚ㍑ศᯒ㸪⏕ែ Ꮫⓗẚ㍑ศᯒࢆ⾜࠺ࡇࡼࡗ࡚㸪⮬↛㑅ᢥࡼࡿ ྍረⓗ⾲⌧ᆺࢹࢨࣥࡢ㐍ࣉࣟࢭࢫࢆ᫂ࡽࡍ ࡿ㸬 㸺ᨭࡢෆᐜ㸼 ᮏ ⏦ ㄳ ㄢ 㢟 ࡛ ࡣ 㸪 ᮏ ✀ ࢆ ᑐ ㇟ ࡋ ࡓ RAD (restriction-site-associated DNA)-seq ࡼࡿつᶍ ࡞㞟ᅋከᆺゎᯒࢆ⾜࠺ࡇࢆᕼᮃࡋࡓ㸬ᚑ᮶㸪㠀ࣔ ࢹࣝ⏕≀ࡢ㑇ఏከᆺゎᯒࡣᩘ㑇ఏᏊᗙ㝈ᐃࡉࢀ࡚ ࠸ࡓࡀࠊሷᇶ㓄ิゎㄞᢏ⾡ࡢⓎ㐩ࡼࡾ㸪RAD-seq ࡢࡼ࠺࡞ࢤࣀ࣒࣡ࢻ࡛ࡢ㑇ఏከᆺゎᯒࡀྍ⬟࡞ ࡗࡓ㸬ඛ⾜◊✲ࡽ࢚ࢰࢧࣥࢩࣙ࢘࢘࢜ࡢ⿕㣗⪅ ࡼࡗ࡚ㄏᑟࡉࢀࡿྍረⓗኚࡣ㸪ᆅᇦ㞟ᅋࡼࡗ࡚ ␗࡞ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸬ࡋࡋ㸪ࡑࡢ㑇ఏⓗ⫼ ᬒࡘ࠸࡚ࡣ mtDNA ᩿∦ࢆ⏝࠸ࡓゎᯒࡀሗ࿌ࡉࢀ ࡚࠸ࡿࡢࡳ࡛㸪ࡇࡢከᵝᛶࡀࡢࡼ࠺ࡋ࡚⋓ᚓࡉ ࢀ㸪㐍ࡋ࡚ࡁࡓࡣᮍࡔ᫂࡞Ⅼࡀከ࠸㸬ࡑࡢ㐣 ⛬ࢆ᫂ࡽࡍࡿࡓࡵ RAD-seq ゎᯒࡼࡗ࡚ᚓࡽ ࢀࡓሗࢆά⏝ࡍࡿ㸬ᮏ◊✲࡛ࡣ㸪ඛ⾜◊✲࡛ࡢ mtDNA ࣁࣉࣟࢱࣉࡢศᕸࢆ⪃៖ࡋ㸪≉≉ᚩⓗ ࡞⾲⌧ᆺࢆࡋࡵࡍあ ᆅᇦ㞟ᅋࡑࡢࡢᆅᇦ㞟ᅋ ࢆẚ㍑ࡍࡿࡓࡵ㸪ᾏ㐨ෆࡢ㸳ࡘࡢᆅᇦ࡛೫ࡾ࡞ࡃ ༸ࡢ᥇㞟ࢆ࠾ࡇ࡞ࡗࡓ㸬ྛ㞟ᅋࡘ࠸࡚ 20 ಶయ㸪ྜ ィ 100 ಶయࡽࢤࣀ࣒ DNA ࢆᢳฟࡋࡓࡢࡕࠊࢤࣀ ࣒ DNA ࢆ EcoRI, BglII ࡢ㸰ࡘࡢไ㝈㓝⣲࡛ษ᩿ࡋ ࡓ㸬ษ᩿ࡉࢀࡓࢤࣀ࣒᩿∦ࢲࣉ࣮ࢱ࣮㓄ิࡢ ຍ࡞ࡢ๓ฎ⌮ࢆ࠾ࡇ࡞࠸ࠊHiseq ࡛ 4 ࣮ࣞࣥศࡢ ࣮࣌ࢻ࢚ࣥࢻࢩ࣮ࢡ࢚ࣥࢩࣥࢢࢆ࠾ࡇ࡞ࡗࡓ㸬 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ࢩ ࣮ ࢡ ࢚ ࣥ ࢩ ࣥ ࢢ ࡢ ⤖ ᯝ 㸪 366,238,834 ࣜ ࣮ ࢻ 㸦3662388 ࣮ࣜࢻ/sample㸧ࡢ࣮ࣜࢻࡀᚓࡽࢀࡓ㸬 ࡍ࡛ྛᆅᇦ㞟ᅋࡢ⾲⌧ᆺࡢྍረⓗኚࡢሗࡣྲྀ ᚓࡋ࡚࠸ࡿࡢ࡛㸪ᚋࡇࢀࡽࡢ⾲⌧ᆺሗᅇࡢ ᨭࡼࡗ࡚ᚓࡽࢀࡓ㑇ఏሗࢆ⤫ྜⓗゎᯒࡋ㸪 ⬨᳝ື≀࠾ࡅࡿྍረᛶࡢ㐍ᶵᵓࡢ㑇ఏⓗᇶ┙ ㏕ࡿணᐃ࡛࠶ࡿ㸬 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ᚋࠊRAD-seq ゎᯒࡢ⤖ᯝᚓࡽࢀࡓ㑇ఏሗࢆά⏝ ࡍࡿࡇ࡛㸪௨ୗࡢࡼ࠺࡞᪂ࡓ࡞◊✲ࡀᒎ㛤ࡉࢀࡿ ࡇࡀᮇᚅࡉࢀࡿ㸬 1) ࢚ࢰࢧࣥࢩࣙ࢘࢘࢜ࡢ⾲⌧ᆺྍረᛶ㛵㐃ࡋࡓ 㞟ᅋ㑇ఏᵓ㐀ゎᯒ㸸 RAD-seq ࡼࡾᚓࡽࢀࡓࢹ࣮ࢱ ࡽ㞟ᅋ㛫ࡢ㛵ಀࡑࡢ㐍ྐࢆ᥎ᐃࡍࡿ㸬あ 㞟 ᅋࢆ୰ᚰࡋࡓᆅᇦ㞟ᅋࡣ㸪ࡢ㞟ᅋ␗࡞ࡿྍረ ⓗኚࢆ♧ࡍࡀ㸪ࡇࡢࡼ࠺࡞㐍ࡀ⮬↛㑅ᢥࡢ⤖ᯝ ⏕ࡌࡓࡢ㸪♽ඛ࠾࠸࡚㞟ᅋࡢ࣎ࢺࣝࢿࢵࢡࢆ⤒ 㦂ࡋ㸪୰❧࡞ኚ␗ࡀ✚ࡋࡓ⤖ᯝ࡞ࡢࡣ᫂ࡽ࡛ ࡣ࡞࠸㸬ࡲࡓ㸪⌧ᅾ㸪ᆅᇦ㞟ᅋ㛫࡛ࡢࡼ࠺࡞㑇ఏ ⓗὶࡀ࠶ࡿࡶ࡛᫂࠶ࡿ㸬RAD-seq ࡼࡗ࡚⏘ ⏕ࡉࢀࡿከᆺࢹ࣮ࢱࢆά⏝ࡍࡿࡇ࡛㸪ࡇࡢࡼ࠺࡞ ၥ㢟ࣉ࣮ࣟࢳࡍࡿ㸬⬨᳝ື≀ࡢ⾲⌧ᆺྍረᛶࡢ 㑇ఏⓗ⫼ᬒࡘ࠸࡚ࡣ㸪ඛ⾜◊✲ࡀஈࡋ࠸ࡓࡵ㸪ᮏ ㄢ㢟ࡢ⤖ᯝࡽࡑࡢᴫせࡀ᫂ࡽ࡞ࢀࡤࠊࡢ◊ ✲⪅᭷┈࡞ሗࢆᥦ౪࡛ࡁࡿᮇᚅࡉࢀࡿ㸬 㸰) ⾲⌧ᆺྍረᛶࡢ㐍ࢆᘬࡁ㉳ࡇࡍ㈐௵㑇ఏᏊࡢ ᥎ᐃ㸸 ከᆺゎᯒࡢ⤖ᯝࡽྛ㞟ᅋ࠾࠸࡚⮬↛㑅ᢥ ࢆཷࡅ࡚࠸ࡿᗙ೫ࡾࡀ࠶ࢀࡤ㸪ࡑࡢ㡿ᇦࢆ≉ᐃ ࡍࡿ㸬⮬↛㑅ᢥࢆཷࡅ࡚࠸ࡿᗙࡣ㸪㞟ᅋ㛫ࡢྍረ ᛶࡢ㐪࠸㛵ಀࡋ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿࡢ࡛㸪ࡑࡢ㡿 ᇦᏑᅾࡍࡿ㑇ఏᏊࡢᶵ⬟ࢆ᥎ᐃࡍࡿ㸬ᡃࠎࡣ᪤ ࢚ࢰࢧࣥࢩࣙ࢘࢘࢜ࡢࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ࢹ࣮ࢱ ࢆಖ᭷ࡋ࡚࠸ࡿࡢ࡛㸪㧗࠸㑅ᢥᅽࢆཷࡅ࡚࠸ࡿᗙ ࡀ㑇ఏᏊ㡿ᇦ୍⮴ࡍࢀࡤ㸪ྍረᛶࡢ㐍┦㛵ࡍ ࡿ㑇ఏᏊࢆ㑅ูࡍࡿࡇࡀ࡛ࡁࡿ㸬᭷ຊ࡞ೃ⿵㑇ఏ Ꮚࡀぢࡘࡗࡓሙྜ㸪ᐃ㔞 PCR ࢆ⏝࠸࡚ྍረᛶㄏᑟ ᐇ㝿㑇ఏᏊⓎ⌧ࡀୖ᪼ࡋ࡚࠸ࡿࢆ☜ㄆࡋ㸪 ᚲせ࡛࠶ࢀࡤᶵ⬟㜼ᐖᐇ㦂ࢆ࠾ࡇ࡞࠺㸬 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࣭‽ഛ୰ - 467 - බເᨭㄢ㢟㸸2012㹼2013 ᖺᗘ ࢩࣕࢪࢡࣔᴫせࢤࣀ࣒ࡢゎㄞ㑇ఏᏛⓗᆅᅗࡢྠᵓ⠏ࡼࡿ㝣ୖ᳜≀㐍ࡢゎ᫂ ᇶ┙◊✲㸦㹀㸧 すᒣ ᬛ࣭᫂㔠ἑᏛᏛ㝿⛉Ꮫᐇ㦂ࢭࣥࢱ࣮ࠊᆏᒣ ⱥಇ࣭⚄ᡞᏛᏛ㝔⌮Ꮫ◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ࢩࣕࢪࢡࣔ┠ࡣࠊ㝣ୖ㏆⦕࡞ࢩࣕࢪࢡࣔ⸴㢮ࡢ ࡞࡛ࡶࠊ༙ᩘయ࠾࠸࡚ჾᐁ⤌⧊ศࢆ♧ࡍ」㞧 ࡞యไࢆࡶࡗ࡚࠸ࡿ⸴㢮࡛࠶ࡿࠋࡇࡢࡓࡵࠊ㝣ୖ᳜ ≀ࡢ㸰ಸయ࡛ࡢከ⣽⬊యไࡢ⋓ᚓ㛵ࢃࡿࢤࣀ࣒㐍 ࢆゎ᫂ࡍࡿࡓࡵࡣᯡせ࡞⨨ࢆ༨ࡵࡿࠋ ࢩࣕࢪࢡࣔࡣࢩࣕࢪࢡࣔ┠ࡢ୰࡛ࢤࣀ࣒ࢧࢬࡀ 2.3Gb ẚ㍑ⓗᑠࡉࡃࠊᐇ㦂ᐊෆ࡛⏕ά⎔ࢆ⤖࡛ ࡁࡿࠋࡇࡢࡓࡵࠊࢩࣕࢪࢡࣔࢆࢤࣀ࣒ゎㄞࡢᑐ㇟ 㑅ᐃࡋࠊࢩࣕࢪࢡࣔࡢࢤࣀ࣒ゎㄞࢆ┠ᣦࡍࡇࡋ ࡓࠋࡲࡓࠊࢩࣕࢪࢡࣔࡣ⏕ែⓗ␗࡞ࡿࢽࢵࢳ࢙ ࢆࡶࡘ✀ෆከᆺࡀ࠶ࡾࠊࡇࡢ㛫ࡢ㓄ࡀྍ⬟࡛࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 ࣭ࢩࣕࢪࢡࣔ S276 ᰴ㸦ὸỈᆺ㸧 ࡢࣥࢧ࣮ࢺ㛗 250bp ࡢ PE ࣛࣈ࣮ࣛࣜࡢㄪᩚ୧➃ࡽ 150 ሷᇶ (PE150)ࡢࢩ࣮ࢣࣥࢩࣥࢢ 5 ࣮ࣞࣥ ࣭ྛ✀ࡢࡁࡉࡢ࣓ࢺ࣌ࣛࣈ࣮ࣛࣜࡢࢩ࣮ࢣ ࣥࢩࣥࢢ PE100 2 ࣮ࣞࣥ PE150㸵࣮ࣞࣥ ࣭ࢩࣕࢪࢡࣔ S277 ᰴ(῝Ỉᆺ)ࡢࣥࢧ࣮ࢺ㛗 250bp ࡢ PE ࣛࣈ࣮ࣛࣜࡢࢩ࣮ࢣࣥࢩࣥࢢ PE150㸯࣮ࣞ ࣥ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᨭࡼࡗ࡚ᚓࡽࢀࡓ S276 ᰴࡢ࣮ࣜࢻࢹ࣮ࢱࢆ ࢭࣥࣈࣝࡋ࡚ࠊScaffold_N50 㛗 2Mb ࢆ㉺࠼ࡿࢭ ࣥࣈ࣮ࣜࢆᚓࡓࠋ ࡇࡢࢭࣥࣈ࣮ࣜ S277 ᰴࡢ㓄ิࢆ࣐ࢵࣆࣥࢢ ࡋࡓࡇࢁࠊmap ࡉࢀࡿ࣮ࣜࢻࡀⴭࡋࡃప࠸ scaffold ࡀྠᐃࡉࢀࠊ࠾ࡑࡽࡃࠊS276 ᰴࡢᇵ㣴ඹᏑࡋ࡚࠸ ࡓࣂࢡࢸࣜࡢ DNA ࡢΰධ࡛࠶ࡿ᥎ᐃࡋࡓࠋࣂࢡ ࢸࣜࡢΰධ᥎ᐃࡉࢀࡿ㓄ิࢆ㝖࠸࡚ᚓࡽࢀࡓࢩ ࣕࢪࢡࣔࡢ㓄ิࡣࠊ11,808 scaffold ィ 1.75Gb, N50 2.26Mb ࡛ࠊ90%ࡀ 550kb ௨ୖࡢ scaffold ྵࡲࢀ ࡚࠸ࡓࠋ ௨๓ᚓ࡚࠸ࡓ RNA-seq ࡢࢹ࣮ࢱཬࡧࠊࢩࣟࢾ ࢼࢬࢼࠊࣄ࣓ࢶࣜ࢞ࢿࢦࢣࡢ࣑ࣀ㓟㓄ิࡢ㢮ఝ ᛶሗࢆ⏝ࡋࠊAugustus ࡼࡗ࡚ࡇࡢࢭࣥࣈࣜ ࣮ୖࡢ㑇ఏᏊᵓ㐀ண ࢆ⾜࠸ࠊ35883 㑇ఏᏊᗙ 36877 ㌿⏘≀ࡢ㑇ఏᏊࣔࢹࣝࢆᚓࡓࠋ ࡇࡢࢤࣀ࣒ࢭࣥࣈ࣮ࣜࣀࢸ࣮ࢩࣙࣥࡢホ౯ ࡋ࡚ CEGMA ࡼࡗ࡚㠀ᖖಖᏑⓗ࡞ 248 ࡢ㑇ఏ Ꮚࢭࢵࢺࡢࢀࡔࡅࡀࢤࣀ࣒ୖ࠶ࡿ࠸ࡣண㌿⏘ ≀㓄ิୖ࡛ぢࡘࡅࡽࢀࡿࢆㄪᰝࡋࡓࠋࡑࡢ⤖ᯝࠊ ࢤࣀ࣒࡛ࡣ 57%complete, 76% partial ࡛ぢࡘࡾࠊ ㌿⏘≀࡛ࡣ 87.5% complete, 91.9% partial ࡛Ⓨぢ ࡉࢀࡓࠋࢤࣀ࣒㓄ิࢆ┤᥋⏝࠸ࡿゎᯒ್࡛ࡀపࡃ࡞ ࡿࡢࡣࠊࣥࢺ࣭࢚ࣟࣥࢡࢯࣥࢆ CEGMA ࡀṇࡋࡃ ㆑ูฟ᮶࡞࠸ࡀ࠶ࡿࡢᑐࡋ࡚ࠊ㑇ఏᏊᵓ㐀ண ࡼࡗ࡚ᚓࡽࢀࡓ㌿⏘≀㓄ิ࡛ࡣ࢚ࢡࢯࣥࡀ㐺ษ 㐃⥆ࡋ࡚ᚓࡽࢀ࡚࠸ࡿࡓࡵ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋ ࡇ࠺ࡋ࡚ᚓࡽࢀࡓ㑇ఏᏊࣔࢹࣝࢆㄪࡿࠊ㝣ୖ ᳜≀ࡢⓎ⏕ࢆไᚚࡍࡿ㑇ఏᏊࡢከࡃࡘ࠸࡚࣍ࣔࣟ ࢢࡀぢࡘࡿࡀ☜ㄆฟ᮶ࡓࠋࡲࡓࠊࣄ࣓ࢶࣜ࢞ࢿ ࢦࢣࡢࢤࣀ࣒ࡣぢࡘࡽ࡞ࡗࡓ㑇ఏᏊࡢ࣍ࣔࣟ ࢢࡶ࠶ࡗࡓࠋ 㑇ఏᏛⓗᆅᅗࢆᵓ⠏ࡍࡿࡣࠊ㓄ศ㞳㞟ᅋࡀᚲ せ࡛࠶ࡿࠋࡇࡇ࡛ࠊࢩࣕࢪࢡࣔࡣ⮬Ṫྍ⬟࡛࠶ࡿࡓ ࡵࠊእ㓄⏤᮶ࡢᏊᏞ⮬ṪࡼࡿᏊᏞࢆ㆑ูࡍࡿ ᚲせࡀ࠶ࡿࠋ῝Ỉᆺࡢ S277 ᰴࡢ࣮ࣜࢻࢆ S276 ࡢ ࢭࣥࣈ࣮࣐ࣜࢵࣆࣥࢢࡋከᆺࢆᢳฟࡋࡓ୰ࡽࠊ ከᆺࢧࢺࡀ BamHI ࡢㄆ㆑㓄ิࡢ᭷↓࡞ࡿ㒊ศ ࢆ㑅ᢤࡋࠊᙜヱ㡿ᇦࢆቑᖜࡍࡿࣉ࣐࣮ࣛࢆタィࡋ ࡓࠋษ᩿ࡉࢀ࡚⏕ࡌࡿ᩿∦㛗ࡀ␗࡞ࡿࡼ࠺」ᩘ㡿 ᇦࢆྠ࣐ࣝࢳࣉࣞࢵࢡࢫ PCR ࡛ቑᖜࡋ࡚ࠊ⤌᭰ ࠼ࢆ᳨ฟฟ᮶ࡿࡼ࠺ࡋࡓࠋࡇࡢࣉ࣐࣮ࣛࢭࢵࢺ ࢆ⏝࠸࡚ࠊඹᇵ㣴ࡢᏊᏞᰴࡢ㑇ఏᏊᆺุᐃࢆ⾜࠺ ࡛ࠊᐇ⏝ⓗ࡞ຠ⋡࡛㓄⏤᮶ᏊᏞᰴࢆྠᐃࡍࡿࡀ ࡛ࡁࠊ⣙㸯㸮㸮ᰴࡢ㓄ᏊᏞᰴࢆྠᐃࡋࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࡇࡢࠊࢩࣕࢪࢡࣔ S276 ᰴࡢࢤࣀ࣒ゎㄞ S277 ᰴࡢ ከᆺ᳨ฟࡼࡾࠊ⮬Ṫ࡛࡞࠸㓄ᰴࡀᚓࡽࢀࡓࡢ࡛ࠊ ࡇࡢศ㞳㞟ᅋࡢከᆺゎᯒࡼࡾ scaffold ࢆᰁⰍయࢫ ࢣ࣮ࣝࡢࢫ࣮ࣃ࣮ࢫ࢟ࣕࢵࣇ࢛࣮ࣝࢻࡃࡳࡓ࡚ࠊ ࢩࣥࢸࢽ࣮ࢆ᳨ウࡍࡿࡀ࡛ࡁࡿࡼ࠺࡞ࡿᮇᚅ ࡉࢀࡿࠋ ࡲࡓࠊ␗࡞ࡿ᮲௳㐺ᛂࡋ࡚␗࡞ࡿᙧ㉁ࢆᣢࡘᰴ 㛫ࡢศ㞳㞟ᅋ࡛࠶ࡿࡢ࡛ࠊᙧ㉁ࢆ ᐃࡋ࡚࣐ࢵࣆࣥ ࢢࡍࡿࡇࡼࡾ㐺ᛂᙧ㉁ࡀࡢࡼ࠺࡞㑇ఏⓗኚ ࡼࡗ࡚㐍ࡋࡓࡢࡢᐇែ㏕ࡿࡀ࡛ࡁࡿᮇ ᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 - 468 - බເᨭㄢ㢟㸸2011 ᖺᗘ ᛶᰁⰍయ㡿ᇦࡢゎᯒᇶ࡙ࡃࠕ㞤㞝ᛶࡢㄌ⏕ࠖ㛵ࡍࡿ㐍⏕≀Ꮫⓗ◊✲ ᇶ┙◊✲㸦㸿㸧 㔝ᓮ ஂ⩏࣭ᮾிᏛᏛ㝔⌮Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ࣓ࢫ࢜ࢫࡢㄌ⏕ࠊࡍ࡞ࢃࡕཎጞⓗ࡞ྠᆺ㓄അࡽ 㞤㞝ࡢ㓄അᏊࢆࡶࡘ⏕≀ࡢ㐍ࡣ᭱ࡶ⯆῝࠸㐍 ⏕≀Ꮫୖࡢၥ㢟࡛࠶ࡿࡀࠊࡑࡢศᏊ㑇ఏᏛⓗᇶ┙ ࡣ࡛᫂࠶ࡗࡓࠋ᭱㏆ᡃࠎࡣ᭷ᛶ⏕Ṫࡢ㐍ࡢࣔࢹ ࣝ⏕≀⩌࡛࠶ࡿ “⩌యᛶ࣎ࣝ࣎ࢵࢡࢫ┠”ࡢࣉࣞ࢜ࢻ ࣜࢼࡽ࢜ࢫ≉␗ⓗ㑇ఏᏊ“OTOKOGI”(PlestMID) ࢆ༢㞳ࡍࡿࡇᡂຌࡋࠊ㞤㞝ࡢᛶྠᆺ㓄അࡢ 㓄ᆺࡢᑐᛂ㛵ಀࢆࡘࡴࡇᡂຌࡋࡓ㸦Nozaki et al. 2006, Cur. Biol.㸧ࠋࡇࡢ㑇ఏᏊࡢⓎぢࡣࡇࢀࡲ ࡛ᅔ㞴ࡉࢀ࡚ࡁࡓ⩌యᛶ࣎ࣝ࣎ࢵࢡࢫ┠ࡢᛶ≉␗ ⓗ㑇ఏᏊࢆྵࡴ“ᛶᰁⰍయ㡿ᇦ”㸦ᛶỴᐃ㑇ఏᏊ㡿ᇦ㸧 ࡢゎᯒ(Ferris et al. 1997, PNAS 94: 8634-8639)ࢆ ⌧ᐇⓗྍ⬟࡞ࡶࡢࡋࡓࠋᮏ◊✲࡛ࡣᛶ≉␗ⓗ㑇ఏ Ꮚ࠾ࡼࡧࡑࢀࡽࡀᏑᅾࡍࡿᛶᰁⰍయ㡿ᇦࡢศᏊ࣭ࢤ ࣀ࣒࣭⣽⬊ࣞ࣋ࣝࡢẚ㍑⏕≀Ꮫⓗゎᯒࢆ⩌యᛶ࣎ࣝ ࣎ࢵࢡࢫ┠ࡢྠᆺ㓄അࡽ␗ᆺ㓄അ࣭༸⏕Ṫࡢ᭷ᛶ ⏕Ṫࡢྛ㐍ẁ㝵ࡢ⏕≀⩌࡛ᐇࡋࠊ ࠕ㞤㞝ᛶࠖࡀㄌ ⏕ࡋࡓศᏊ⣽⬊Ꮫⓗ࡞ᇶ┙ࢆ᫂ࡽࡍࡿࡇࢆ┠ ⓗࡋ࡚࠸ࡿࠋ 㸺ᨭࡢෆᐜ㸼 2011 ᖺᗘࡣ⥳⸴⩌యᛶ࣎ࣝ࣎ࢵࢡࢫ┠࡛⩌యࡋࡓ ึᮇẁ㝵࡛࠶ࡾࠊྠᆺ㓄അࡢ᭷ᛶ⏕Ṫࡢࢦࢽ࣒࢘ 㸦Gonium pectorale㸧ࡢ┦⿵ⓗ㓄ᆺࡢࣉࣛࢫᰴ ࣐ࢼࢫᰴࡢࢤࣀ࣒ゎᯒࡢᨭࢆᚓࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 ᡃࠎࡣࢦࢽ࣒࢘ࡢᛶᰁⰍయ㡿ᇦࡢࢤࣀ࣒ゎㄞࢆ┠ⓗ ᖹᡂ 23 ᖺᗘࢤࣀ࣒ᨭࢆᚓࡓࠋࡑࡢ⤖ᯝࠊ࢜ࣝ࢞ ࢿࣛࢤࣀ࣒㓄ิ㸦Hamaji et al. 2013, PLoS ONE㸧᰾ࢤࣀ࣒㓄ิ㸦Hanschen et al. ཎ✏୰㸧 ࡀᵓ⠏ࡉࢀࡓࠋࢦࢽ࣒࢘ࡇࢀࡲ࡛ࡢࡢ⩌యᛶ࣎ ࣝ࣎ࢵࢡࢫ┠ࡢ࢜ࣝ࢞ࢿࣛࢤࣀ࣒(Smith and Lee 2010, MBE; Smith et al. 2013, MBE)ࢆẚ㍑ࡍࡿࠊ ⣽⬊ᩘࡢቑຍඹ࢜ࣝ࢞ࢿࣛࢤࣀ࣒ࡢ㠀ࢥ࣮ࢻ㡿 ᇦࡀቑຍࡋ࡚࠸ࡿࡇࡀ⪃࠼ࡽࢀࡓࠋࡇࡢࡼ࠺࡞㠀 ࢥ࣮ࢻ㡿ᇦࡢቑຍ⌧㇟ࡣ㸯ಶయ࠶ࡓࡾࡢ⣽⬊ᩘࡀቑ ຍࡍࡿࡇ㸻㞟ᅋࢧࢬࡀ⦰ᑠࡍࡿࡇࡀཎᅉ࡛࠶ ࡿࡍࡿ࣑࣮ࣗࢸ࣮ࢩࣙࢼࣝࣁࢨ࣮ࢻㄝ 㸦mutational-hazard hypothesis㸧(Lynch 2007. The origins of genome architecture. Sinauer Assoc.)࡛ ㄝ᫂ࡉࢀࡿࠋ ࡲࡓࠊࡇࢀࡲ࡛⦰㔜ࣉ࣐࣮ࣛἲ࡛ࡣྲྀᚓࡍࡿࡇ ࡢ࡛ࡁ࡞ࡗࡓࢦࢽ࣒࢘㓄അᏊࡢࠕ⼥ྜࠖ㛵ࡍࡿ 㑇ఏᏊ GCS1(HAP2) ࡢ࣍ࣔࣟࢢ㸦GpGCS1㸧ࡢ㓄 ิࡀ᰾ࢤࣀ࣒㓄ิሗࡽ᫂ࡽ࡞ࡗࡓࠋ GpGCS1 ࡣ㠀ᖖ G+C ẚࡢ㧗࠸㓄ิ࡛࠶ࡾࠊࢤ ࣀ࣒ゎᯒࡀ࡞ࡅࢀࡤᚓࡽࢀ࡞࠸ࡶࡢ⪃࠼ࡽࢀࡓ (Kawai-Toyooka et al. 2014, Eukaryotic Cell)ࠋ୍᪉ࠊ ࢦࢽ࣒࢘᰾ࢤࣀ࣒ࢆ᥈⣴ࡋࡓ⤖ᯝࠊࢡ࣑ࣛࢻࣔࢼ ࢫ࣎ࣝ࣎ࢵࢡࢫࡢᛶᰁⰍయ㡿ᇦᏑᅾࡋࡓከࡃࡢ 㑇ఏᏊࡢ࣍ࣔࣟࢢࡀࢦࢽ࣒࢘ࡢᛶᰁⰍయ㡿ᇦ௨እࡢ ᖖᰁⰍయ㡿ᇦ☜ㄆࡉࢀࠊᛶᰁⰍయᖖᰁⰍయ㡿ᇦ 㛫ࡢ㢖⦾࡞㑇ఏᏊࡢ㐍ⓗ⛣ືࡀ♧၀ࡉࢀࡓ 㸦Hamaji et al. ᢞ✏୰㸧 ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ࢦࢽ࣒࢘ࡢᛶᰁⰍయ㡿ᇦࡣࢡ࣑ࣛࢻࣔࢼࢫẚ㍑ࡍ ࡿ㠀ࢥ࣮ࢻ㡿ᇦࡀቑࡍࡿࡀࠊ࣎ࣝ࣎ࢵࢡࢫࡢᛶ ᰁⰍయ㡿ᇦ࡛᫂ࡽ࡞ࡗࡓ᪂つ࣓ࢫ࣭࢜ࢫ≉␗ⓗ 㑇ఏᏊ㸦Ferris et al. 2010, Science㸧ࡢ࣍ࣔࣟࢢࡣ ࢦࢽ࣒࢘ࡢࢤࣀ᳨࣒࡛ࣞ࣋ࣝฟࡉࢀ࡞ࡗࡓࠋᚑ ࡗ࡚ࠊ࣎ࣝ࣎ࢵࢡࢫࡢ᪂つ࣓ࢫ࣭࢜ࢫ≉␗ⓗ㑇ఏᏊ ࡢ㉳※ࢆ᫂ࡽࡍࡿࡣࠊࡼࡾ࣎ࣝ࣎ࢵࢡࢫ㏆ ⦕࡞⩌యᛶ࣎ࣝ࣎ࢵࢡࢫ┠ࡢྠᵝࢤࣀ࣒ゎᯒࡀ ᚲせ࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ࢤࣀ࣒ࢻࣛࣇࢺ㓄ิࡢ⤖ᯝࠊࡇࢀࡲ࡛ 2 ᖺ㛫 ࡅ࡚⦰㔜ࣉ࣐࣮ࣛἲ࡛᥈⣴ࡋ࡚ᡂຌ࡛࠶ࡗࡓ㓄 അᏊ⼥ྜ㛵㐃ࡍࡿ㑇ఏᏊ GCS1 ࣍ࣔࣟࢢ GpGCS1 ࢆᚓࡿࡀ࡛ࡁࡓࠋᚓࡽࢀࡓ GpGCS1 ࡢ cDNA 㓄 ิࡣ GC ྵ㔞ࡀ 72%࡛࠶ࡾࠊ࡚ࡶᚑ᮶ࡢ PCR ἲ ࡛ࡣ᥈⣴࡛ࡁ࡞࠸ࡇ᪂ᆺࢩ࣮ࢣࣥࢧ࣮ࡢጾຊࢆ ᨵࡵ࡚⌮ゎ࡛ࡁࡓࠋ᪂ᆺࢩ࣮ࢣࣥࢧ࣮࡛ࢻࣛࣇࢺ㓄 ิࢆᚓ࡚ᕼᮃࡢ㑇ఏᏊࢆᚓࡿ࠸࠺᪂ࡓ࡞᪉ἲㄽࢆ య㦂࡛ࡁࠊᚋࡇࡢࡼ࠺࡞᪉ἲࢆ㥑ࡋ࡚ࠊ㠀ࣔࢹ ࣝ⏕≀࡛ࡶ༑ศ◊✲ࡀ࡛ࡁࡿ࠸࠺ᒎᮃࡀᣅࡅࡓࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 Hamaji, T. et al., PLoS ONE 8, e57177 (2013) Kawai-Toyooka, H. et al., Eukaryotic Cell 13, 648-656 (2014) - 469 - බເᨭㄢ㢟㸸2012 ᖺᗘࠊ2014 ᖺᗘ ᛶᰁⰍయ㡿ᇦࡢẚ㍑ゎᯒᇶ࡙ࡃࠕ㞤㞝ᛶࡢㄌ⏕ࠖࡢ㐍⏕≀Ꮫⓗ◊✲ ᇶ┙◊✲㸦㸿㸧 㔝ᓮ ஂ⩏࣭ᮾிᏛᏛ㝔⌮Ꮫ⣔◊✲⛉ 㸺⫼ᬒ࣭┠ⓗ㸼 ࣓ࢫ࢜ࢫࡢࡣࡌࡲࡾࡣᡃࠎே㢮ࡢ㛗ᖺࡢ㢟࡛࠶ ࡾࠊ⏕≀Ꮫⓗࡣྠᆺ㓄അࡽ༸⢭Ꮚࢆࡶࡘ⏕≀ ࡢ㐍࡛࠶ࡿࠋᡃࠎࡣᛶ㐍ࡢࣔࢹࣝ⏕≀⩌ࠕ⩌ యᛶ࣎ࣝ࣎ࢵࢡࢫ┠ࠖࡢࣉࣞ࢜ࢻࣜࢼࡽ࢜ࢫ≉␗ ⓗ㑇ఏᏊ“OTOKOGI”ࢆ༢㞳ࡍࡿࡇᡂຌࡋࠊ㞤㞝 ࡢᛶྠᆺ㓄അࡢ㓄ᆺࡢᑐᛂ㛵ಀࢆึࡵ࡚᫂ࡽ ࡋࡓ㸦Nozaki et al. 2006, Curr. Biol.16: R1018㸧ࠋ ᮏ㑇ఏᏊࡢⓎぢࡣࡇࢀࡲ࡛ᅔ㞴ࡉࢀ࡚ࡁࡓ⩌యᛶ ࣎ࣝ࣎ࢵࢡࢫ┠ࡢᛶỴᐃ㑇ఏᏊࡀ⨨ࡍࡿࠕᛶᰁⰍ య㡿ᇦ (MT)ࠖࡢࢤࣀ࣒ゎᯒࡢᐇ⌧ᛶࢆ♧၀ࡋࡓࠋ᭱ ㏆ᡃࠎࡣ༸⏕Ṫ࣎ࣝ࣎ࢵࢡࢫࡢ㞤㞝 MT ࢆゎㄞࡋࠊ ྠᆺ㓄അࢡ࣑ࣛࢻࣔࢼࢫẚ㍑ࡋࡓ(Ferris et al. 2010, Science 328: 351)ࠋࡑࡢ⤖ᯝࠊMT ࡣᣑ࣭୧ ᴟࡉࢀࠊ13 ಶࡢ㞤㞝≉␗ⓗ࡞᪂つ㑇ఏᏊࢆⓎぢ ࡋࡓࠋࡋࡋࠊ୧ᴟ➃࡞⏕≀ࡢ MT ࡢẚ㍑࡛࠶ࡾࠊ 㐍୰㛫ẁ㝵ࡢ◊✲ࡢᚲせᛶࡀ㆟ㄽࡉࢀࡓ 㸦Charlesworth & Charlesworth 2010, Curr. Biol. 20:R519㸧ࠋᮏ◊✲ࡢ┠ⓗࡣࠊ⩌యᛶ࣎ࣝ࣎ࢵࢡࢫ┠ ࡢྠᆺ㓄അࡽ␗ᆺ㓄അ࣭༸⏕Ṫࡢྛ㐍ẁ㝵ࡢ⏕ ≀࡛ MT ᇦࢆ᫂ࡽࡋࠊẚ㍑ࢤࣀ࣒࣭㐍⏕≀ Ꮫⓗゎᯒࢆᐇࡋࠊ ࠕ㞤㞝ᛶࡢ㉳※ࠖ୪ࡧࠕ࣓ࢫࡽ ࡋࡉࠖࠕ࢜ࢫࡽࡋࡉࠖࡢ⋓ᚓ㐍ࡢ┤᥋ⓗ࡞ཎᅉ ࡞ࡗࡓศᏊ㑇ఏᏛⓗ࡞ᇶ┙ࢆ᫂ࡽࡍࡿࡇ࡛ ࠶ࡿࠋ 㸺ᨭࡢෆᐜ㸼 2011 ᖺᗘᨭࢆཷࡅࡓࢦࢽ࣒࢘ࡼࡾ࣎ࣝ࣎ࢵࢡࢫ ࡼ ࡾ ㏆ ⦕ ࡞ ྠ ᆺ 㓄 അ ࡢ ࣖ ࣐ ࢠ ࢩ ࢚ ࣛ 㸦 Yamagishiella 㸧ࠊ 㞤 㞝 ᛶ ࢆ ࡶ ࡘ ࣘ ࣮ ࢻ ࣜ ࢼ 㸦Eudorina㸧ࡣ㞤㞝ᛶࡢ㐍ࡢゎ᫂ࡢࡓࡵࡢẚ㍑◊ ✲ 㔜 せ ࡛ ࠶ ࡿ ࠋ ࡇ ࡢ ࡓ ࡵ Eudorina sp. Yamagishiella unicocca ࡢ୧ᛶࡢ᰾ࢤࣀ࣒ࢆᵓ⠏ ࡋࡓࠋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 Eudorina sp. Yamagishiella unicocca ࡢ୧ᛶࡢ ᰾ࢤࣀ࣒ࢹ࣮ࢱࡀᚓࡽࢀࡓࠋࡑࡢ⤖ᯝࠊࢡ࣑ࣛࢻ ࣔࢼࢫࡢ࣐ࢼࢫ≉␗ⓗ㑇ఏᏊ࡛࠶ࡿ MID ࡢ࣍ࣔ ࣟ ࢢ ࡀ Eudorina sp. ࢜ ࢫ ᰴ Yamagishiella unicocca ࣐ࢼࢫᰴ࡛ࡑࢀࡒࢀᛶ≉␗ⓗ᳨ฟࡉ ࢀࡓࠋࡲࡓࠊࢡ࣑ࣛࢻࣔࢼࢫࡢࣉࣛࢫ≉␗ⓗ㓄അᏊ ᥋╔㑇ఏᏊ࡛࠶ࡿ FUS1 ࣍ࣔࣟࢢࡀ Eudorina sp. ࣓ࢫᰴ Yamagishiella unicocca ࣉࣛࢫᰴ࡛ࡑࢀࡒ ࢀᛶ≉␗ⓗ᳨ฟࡉࢀࡓࠋ௨ୖ⤖ᯝࡣࡼࡾ⣔⤫ⓗ ᇶ㒊⨨ࡍࡿࢦࢽ࣒࢘㸦Hamaji et al. ᢞ✏୰㸧 ྠࡌ࡛࠶ࡗࡓࠋ༸⏕Ṫࡢ࣎ࣝ࣎ࢵࢡࢫ࡛ࡣ FUS1 ࣍ࣔࣟࢢࡀㄆࡵࡽࢀ࡞࠸ࡀ㸦 Ferris et al. 2010, Science㸧ࠊ␗ᆺ㓄അࡢ Eudorina sp. ㄆࡵࡽࢀࡿࡢ ࡛ࠊᮏ㑇ఏᏊࡢᾘኻࡣ␗ᆺ㓄അ௨㝆ࡢẁ㝵࡛㓄അ Ꮚࡢ᥋╔ᵝᘧࡢ㐍కࡗ࡚㉳ࡁࡓࡇࡀ᥎ ࡉࢀ ࡓ㸦㔝ᓮࡽ 2014 ᖺ᪂Ꮫ⾡㡿ᇦ◊✲ࠕࢤࣀ࣒ᨭࠖᣑ ⌜㆟㸧 ࠋ୍᪉ࠊࢡ࣑ࣛࢻࣔࢼࢫࢦࢽ࣒࢘ࡢྠᆺ 㓄അࡢ୧✀࡛࣐ࢼࢫ㓄ᆺ㸦MID ࢆࡶࡘ㸧ࡔࡅ ㄆࡵࡽࢀ㸦Hamaji et al. 2009, J. Phycol.㸧ࠊ༸⏕Ṫ ࡢ࣎ࣝ࣎ࢵࢡࢫ࡛ࡣ㞝ࡔࡅഇ㑇ఏᏊࡋ࡚ㄆࡵࡽ ࢀ ࡿ 㸦 Ferris et al. 2010 㸧 MTD1 ࣍ ࣔ ࣟ ࢢ ࡣ Eudorina sp. Yamagishiella unicocca ࡢࡑࢀࡒࢀ ࡢ୧ᛶࡢࢤࣀ࣒ㄆࡵࡽࢀࡓࠋࡲࡓࠊࡇࢀࡲ࡛ࢡ ࣑ࣛࢻࣔࢼࢫࡸࢦࢽ࣒࢘ࡑࡢ࣍ࣔࣟࢢࡢㄆࡵࡽࢀ ࡞ࡗࡓ࣎ࣝ࣎ࢵࢡࢫࡢ࣓ࢫ≉␗ⓗ㑇ఏᏊࡢ࣍ࣔࣟ ࢢࡀ Yamagishiella unicocca ࡢࢤࣀ࣒ࡣㄆࡵࡽࢀ ࡞ࡗࡓࡀࠊ2 㑇ఏᏊࡀ␗ᆺ㓄അࡢ Eudorina sp.ࡢ ୧ᛶࡢࢤࣀ᳨࣒ฟࡉࢀࡓࠋᚑࡗ࡚ࠊᛶ≉␗ⓗ㑇ఏ Ꮚࡣ㐍ẁ㝵࣭⣔⤫ࡼࡗ࡚ࡣ୧ᛶᏑᅾࡍࡿሙྜ ࡶ࠶ࡿࡇࡀ᫂ࡽ࡞ࡗࡓ㸦ᮍⓎ⾲㸧ࠋ Eudorina sp. Yamagishiella unicocca ࡢⰍ⣲య ࢤࣀ࣒ࡀỴᐃࡋࡓࠋඹ⎔≧ࢤࣀ࣒࡛࠶ࡗࡓࡀࠊࡢ ࡶࡢࡀࡁࡃࠊྠࡌ⣽⬊ᩘࡢ Eudorina sp. ࡢࡶࡢ ࡼࡾࡶࣜࣆ࣮ࢺࡀከࡃ᳨ฟࡉࢀࡓࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 ⩌యᛶ࣎ࣜ࣎ࢵࢡࢫ┠࡛⣽⬊ᩘࡣྠࡌࡔࡀ᭷ᛶ⏕ Ṫ ࡀ ␗ ᆺ 㓄 അ ࡢ Eudorina sp. ྠ ᆺ 㓄 അ ࡢ Yamagishiella unicocca ࡢࢤࣀ࣒ẚ㍑ࡽࡇࢀࡲ࡛ ࡑࡢ㉳※ࡀࡃ࡛᫂࠶ࡗࡓ࣎ࣝ࣎ࢵࢡࢫ࣓ࢫ≉␗ ⓗࡢ࣍ࣔࣟࢢࡀ୧✀ࡢ୧ᛶ࡛᫂ࡽ࡞ࡗࡓࠋࡇࡢ ࡇࡣ୧ᛶࡢࢤࣀ࣒࡛᪂つ㑇ఏᏊࡀ㐍ࡋࠊ༸⏕Ṫ ࡢ࣎ࣝ࣎ࢵࢡࢫ㐍ࡍࡿ௨๓∦᪉ࡢᛶࡢࢤࣀ࣒ ೫ᅾࡍࡿ㐍ࡀ᥎ ࡉࢀࡓࠋ᭦୧✀ࡢࢤࣀ࣒ゎ ᯒࢆ㐍ᒎࡉࡏࠊᛶᰁⰍ㡿ᇦࢆ᫂ࡽࡋࠊᛶᰁⰍయ ࡢ㐍ඹ㆟ㄽࡍࡿᚲせࡀ࠶ࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ࡞ࡋ - 470 - බເᨭㄢ㢟㸸2015 ᖺᗘ ࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ 1HR ᛶᰁⰍయࢆ⏝࠸ࡓ < ᰁⰍయࡢ㏥㐣⛬ࡢゎ᫂ ᣮᡓⓗⴌⱆ◊✲ 㔝⃝ ᫀᩥ࣭ᅜ❧㑇ఏᏛ◊✲ᡤ ⏕ሗ◊✲ࢭࣥࢱ࣮ 㸺⫼ᬒ࣭┠ⓗ㸼 ᛶᰁⰍయࡀ⏕ࡌࡿࠊ㏻ᖖ Y ᰁⰍయࡣከࡃࡢ㑇ఏ Ꮚࢆኻ࠸㏥ࡋ࡚࠸ࡃࠋࡋࡋࠊࢇࡢ⏕≀ࡢ Y ᰁⰍయࡣ㉳※ࡀ㠀ᖖྂࡃࠊ᪤㒊ศࡀ࣊ࢸࣟࢡ ࣐ࣟࢳࣥࡋ࡚࠸ࡿࡓࡵࠊࡑࡢ㏥㐣⛬ࢆ᫂ࡽ ࡍࡿࡇࡀ࡛ࡁ࡞࠸ࠋࡑࡇ࡛ᮏ◊✲࡛ࡣࠊ᭱㏆ᛶᰁ Ⰽయࡀ⊂❧⏕ࡌࡓ 3 ✀ࡢࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ࢆ⏝࠸ ࡚ࠊY ᰁⰍయࡢ㏥㐣⛬ࢆゎ᫂ࡍࡿࡇࢆ┠ⓗࡍ ࡿࠋᮏ◊✲ࢆ㐙ࡉࡏࠊY ᰁⰍయࡽࡢ㑇ఏᏊᾘኻ 㐣⛬ࢆ᫂ࡽࡍࡿࠋࡲࡓࠊ㌿⛣ᅉᏊࡢ✚ࡼࡿ Y ᰁⰍయࡢ࣊ࢸࣟࢡ࣐ࣟࢳࣥࡢ㐣⛬ࢆゎ᫂ࡍࡿࠋ 㸺ᨭࡢෆᐜ㸼 ᭱㏆ᛶᰁⰍయࢆ⋓ᚓࡋࡓ 3 ✀ࡢࢩࣙ࢘ࢪࣙ࢘ࣂ࢚ ࡢ ࠺ ࡕ ࡢ 2 ✀ 㸦 Drosophila albomicans D. americana㸧ࡢ࣓ࢫࡘ࠸࡚ࠊPacBio ࢩ࣮ࢣࣥࢫ㸦⣙ 100㸧ࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓ㸦୍㒊⌧ᅾ㐍⾜୰㸧ࠋࡲ ࡓࠊࡇࡢ 2 ✀ࡢ࢜ࢫ࣓ࢫࡢ୧᪉ࡘ࠸࡚ Illumina ࢩ࣮ࢣࣥࢫ㸦⣙ 50㸧ࢆ⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋࡲࡓࠊ PacBio ࢩ࣮ࢣࣥࢫࢹ࣮ࢱࢆ⏝࠸ࡓࢤࣀ࣒ࢭࣥࣈࣝ ࡶ⾜࡞ࡗ࡚࠸ࡓࡔ࠸ࡓ㸦⌧ᅾࢭࣥࣈࣝ⢭ᗘ᳨ド ࢆྠ㐍⾜୰㸧 ࠋ 㸺ᨭࡼࡗ࡚㛤ࢀࡓ᪂ࡓ࡞ᒎᮃ㸼 PacBio ࢩ࣮ࢣࣥࢫࡼࡿࢤࣀ࣒㓄ิỴᐃࢆ⾜ࡗࡓ ࡇ࡛ࠊࡇࢀࡲ࡛ゎᯒࡍࡿࡇࡀᅔ㞴࡛࠶ࡗࡓ㑇ఏ Ꮚእ㡿ᇦࠊ≉㓄ิࢆྵࡴ㡿ᇦࡶ↔Ⅼࢆᙜ࡚ ࡿࡇࡀྍ⬟࡞ࡗࡓࠋࡇࢀࡼࡾࠊᛶᰁⰍయ క࠸ࠊࡢࡼ࠺㓄ิࡀ✚ࡋ࡚࠸ࡃࡢࠊࡲ ࡓ✚ࡋࡓ㓄ิࡀࡢࡼ࠺㑇ఏᏊ㡿ᇦࡢ㏥ 㛵ࡋ࡚࠸ࡿࡢࢆゎ᫂ࡍࡿ㊊ࡾ࡞ࡗࡓ ࠸࠼ࡿࠋ ࡲࡓࠊᮏᨭࢆཷࡅࡓࡇ࡛ࠊ◊✲ண⟬ࢆᙜึィ ⏬ྵࡲࢀ࡚࠸࡞ࡗࡓ RNA-seq ⏝ࡍࡿࡇ ࡀྍ⬟࡞ࡗࡓࠋࡇࢀࡼࡾࠊࢤࣀ࣒㓄ิࡽࡔࡅ ࡛ࡣᅔ㞴࡞㑇ఏᏊࡢࠕᶵ⬟ᛶ㸦ࡍ࡞ࢃࡕᶵ⬟㑇ఏᏊ ഇ㑇ఏᏊ㸽㸧 ࠖࡘ࠸࡚ࠊ㑇ఏᏊⓎ⌧ࡢ᭷↓࠸ ࠺᪂ࡓ࡞ሗࢆຍࡍࡿࡇࡀ࡛ࡁࡿࠋࡋࡓࡀࡗ࡚ࠊ ࡼࡾ⢭ᗘࡢ㧗࠸㑇ఏᏊࣀࢸ࣮ࢩࣙࣥࢆ⾜࠺ࡇࡀ ྍ⬟࡞ࡿࡣࡎ࡛࠶ࡿࠋࡋࡓࡀࡗ࡚ࠊY ᰁⰍయࡢ㑇 ఏᏊ㡿ᇦࡢ㏥㐣⛬ࡘ࠸࡚ࠊᙜึィ⏬ࢆୖᅇࡿ▱ ぢࡀᚓࡽࢀࡿࡇࡀᮇᚅࡉࢀࡿࠋ 㸺ᨭᡂᯝබ⾲ࣜࢫࢺ㸼 ヱᙜ࡞ࡋ 㸺ᨭࡼࡗ࡚ᚓࡽࢀࡓ◊✲ᡂᯝ㸼 D. albomicans 㛵ࡋ࡚ࡣࠊ᪤࣓ࢫࡢ DNA ࢆ⏝ ࠸ࡓ PacBio ࢩ࣮ࢣࣥࢫࡀࡋࠊࢭࣥࣈࣝࡲ࡛⤊ ࡋ࡚࠸ࡿࠋ⌧ᅾࠊࢭࣥࣈࣝࡢ⢭ᗘࢆ᳨ド୰࡛࠶ ࡿࡀࠊࢥࣥࢸࢢࡢ N50 ࡀ⣙ 22Mbpࠊ⥲ࢥࣥࢸ ࢢ㛗ࡢ 8 ࡣ 9 ᮏࡢࢥࣥࢸࢢ࡛ᵓᡂࡉࢀࡿࠊ࠸ ࠺⤖ᯝ࡞ࡾࠊࡇࢀࡲ࡛ࡢ Illumina ࢩ࣮ࢣࣥࢫ࡛ࡣ ⪃࠼ࡽࢀ࡞࠸㧗⢭ᗘ࡞ࢤࣀ࣒㓄ิࡀỴᐃ࡛ࡁࡓ ࡢ࡛ࡣ࡞࠸⪃࠼࡚࠸ࡿࠋIllumina ࢩ࣮ࢣࣥࢫ ࡘ࠸࡚ࡣࠊ࢜ࢫ࣓ࢫ୧᪉࡛⾜ࡗ࡚࠸ࡓࡔ࠸ࡓࠋPacBio ࢭࣥࣈࣜࡀḟ➨ࠊ࢜ࢫ࣓ࢫࡢ Illumina ࣮ࣜ ࢻࢆ࣐ࢵࣆࣥࢢࡋࠊ࢝ࣂ࣮ࣞࢪẚࡽᛶᰁⰍయࡢ㏥ ࡢ⛬ᗘࢆ᥎ᐃࡍࡿணᐃ࡛࠶ࡿࠋ D. americana ࡘ࠸࡚ࡣࠊྠ㉁㑇ఏᏊ⣔⤫ࡢ☜❧ ࢆ⾜࠺௨๓ࠊࣁ࢚ࡢ㣫⫱ᡭ㛫ྲྀࡾࠊ࡞࡞ DNA ࡢᢳฟࢆ⾜࠺ࡇࡀฟ᮶࡞ࡗࡓࠋ11 ᭶ධ ࡾࠊࡼ࠺ࡸࡃ DNA ࢆᢳฟࡍࡿࡔࡅࡢࣁ࢚ࡀ㞟ࡲࡾࠊ 12 ᭶ᮎࡲ࡛ DNA ᢳฟࡀࡋࡓࠋPacBio ࢩ࣮ࢣ ࣥࢫࡘ࠸࡚ࡣࠊD. albomicans ྠᵝࡢ⢭ᗘࡢ࣓ࢫ ࢤࣀ࣒㓄ิࡀᚓࡽࢀࡿࡇࢆᮇᚅࡋ࡚࠸ࡿࠋ Illumina ࢩ࣮ࢣࣥࢫࡘ࠸࡚ࡣ᪤࢜ࢫ࣓ࢫ୧᪉ࡢ DNA ࢆ⏝࠸ࡓࢩ࣮ࢣࣥࢫࡀࡋ࡚࠸ࡿࠋPacBio ࢭࣥࣈࣜࡀḟ➨ࠊD. albomicans ྠᵝ࢜ࢫ࣓ࢫ ࡑࢀࡒࢀࡢ࣮ࣜࢻࢆ࣐ࢵࣆࣥࢢࡋࠊ࢝ࣂ࣮ࣞࢪẚ ࡽᛶᰁⰍయࡢ㏥ࡢ⛬ᗘࢆ᥎ᐃࡍࡿࠋ - 471 -
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