Nucleic Acids Research
Vdume 1 2 umber 3 1984
2 ' , 3'-Dldeoxy-3' aminonlldeoside S1-Criphosphataare the terminators of DNA synthesis cntPlyzed
by DNA p o l y m ~
Z.G.Chidgeavadze and R.Sh.Beabealashvilli
National Cardiology Research Center of the USSR Academy of Medical Sciences, 3-rd
Cherepkovskaya str., 15A, Moscow 121522, USSR
A.M.Atrazhev, M.K.Kukhanova, A.V.Azhayev and A.A.Krayevsky
Institute of Molecular Biology of the USSR Academy of Sciences, Vavilov str., 32, Moscow 117984,
USSR
Received 20 October 1983; Accepted 14 December 1983
ABSTRACT
It is shown that 2',3'-dideoxy-3'-aminonucleoside 5'-triphosphates with adenine, guanine, cytosine and thymine bases are
effective inhibitors of DNA polymerase I, calf thymus DNA polymerase o( and rat liver DNA polymerase p
The effect of the
above-mentioned compounds is markedly higher than corresponding
action of the well-known DNA synthesis inhibitors arabinonucleoside 5'-triphosphates and 2',3'-dideoxynucleoside 5'-triphosphates. 2',3'-dideoxy-3'-aminonucleoside 5'-monophosphate residues
incorporate into the 3'-terminus of the primer and terminate the
DNA chain elongation. The possibility of using 2',3'-dideoxy-3'aminonucleoside 5'-triphosphates as terminators for DNA sequencing by the polymerization method is damonstzated.
.
INTRODUCTION
In I969 Atkinson et al. (I) reported that ddNTP served as
inhibitors of DNA synthesis being a competitive agent for dTTP in
the process catalyzed by E.coli DNA polymerase I. ddNTP reacted
with the 3'-nucleotide residue of the primer, incorporated into
the 3'-position of the primer and blocked the chain elongation.
Later, Sanger and coworkers showed (2) that all the four ddNTP ddTTP, ddATP, ddGTP and ddCTP possessed termination properties,
every ddNTP competing with dNTP of the corresponding nature. This
property of ddNTP underlies the sequencing of DNA by the polymerization method (2,3)
At the same time, the application of ddNTP as terminators is
restricted. For DNA sequencing, it implies a comparatively low
efficiency of incorporation into the DNA chain as compared with
dNTP. It is shown, that the reaction rate of ddTTP with primer
(tested on activated DNA and E.coli DNA polymerase I) is 1000
times lower than the rate of dTTP incorporation (I).
Therefore, for DNA sequencing one has to use large excess
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43 IRL Press U m ~Oxford,
,
England.
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