Supplementary Material
1 2 3
4
Fig. S1. PCR amplification to confirm homologous recombination in Nicotiana
benthamiana.
Total RNA that was isolated from the fourth upper leaf infected by pCF93-egfp (lane 1),
pCF93K-egfp (lane 2), pCF157-egfp (lane 3), or pCF157K-egfp (lane 4) was used for PCR.
The PCR involved primers SJ187 and CFCP-R. The red arrow shows the size of wild-type
cucumber fruit mottle mosaic virus (CFMMV), and the blue arrow indicates the size of
CFMMV carrying enhanced green fluorescent protein (EGFP).
1
1
Table S1.
2
Primers used for construction of coat protein deletion mutants.
Restriction
site
(italicized
sequence)
Primer
Sequence
Name of
constructed vector
CF+68-R
5′-ATTACGCGT GTTTAAACCTCGAGAAGCATAATAAGGACAAAAAG-3′
pCF(+)68-egfp
XhoI-PmeIMluI
CF+88-R
5′-ATTCTCGAGACACCAACAGATCATACAAAG-3′
pCF(+)88-egfp
XhoI
CF+100-R
5′-ATTCTCGAGCACCTTGGGCTGACACCAACA-3′
pCF(+)100-egfp
XhoI
CF+127-R
5′-ATTCTCGAGCTTTACCATTTTGCGTTTGCA-3′
pCF(+)127-egfp
XhoI
CF+140-R
5′-ATTCTCGAGTTAACAACCCATTTATGGAGT-3′
pCF(+)140-egfp
XhoI
CF+160-R
5′-ATTCTCGAGTTAACAACCCATTTATGGAGT-3′
pCF(+)160-egfp
XhoI
SJ187
5′-AAAGCTGCCTTTTGTGGTGAT-3′
SJ395
5′-ACCAGAAGTAGAGTAAGACGT-3′
SJ396
5′-ACGTCTTACTCTACTTCTGGT-3′
CFNTR-F
5′-ATTGTTTAAACGGCGTGGTCGCTGCGTTA-3′
CF-3Sac
5′-ATTCTGCAGAAAAGGTTCAACCCTTTTTTG-3′
3
4
5
6
7
8
9
2
CP deletion mutant:
Step I
CP deletion mutant:
Step II
PmeI
SacI
10
Table S2.
11
Primers used for construction of movement protein deletion mutants.
12
3
13
Table S3.
14
Primers used for construction of expression vectors based on cucumber fruit mottle mosaic virus.
15
16
4
17
Table S4.
18
Primers used for real-time quantitative RT-PCR.
Primer
Sequence
Target plant
and/or genes
Product size
GADPH-qRT F
5′-GGAGGAGGGAACAACAAGAGG-3′
N. benthamiana
238 bp
GADPH-qRT R
5′-AGATGCCGTCAGTGCCGA-3′
Glyceraldehyde 3phosphate
dehydrogenase
EGFP-qRT F
5′-AGAACGGCATCAAGGTGAAC-3′
EGFP-qRT R
5′-TGCTCAGGTAGTGGTTGTCG-3′
CFCP-qRT F
5′-GGTTGTTAACGACAGTTGCG-3′
CFCP-qRT R
5′-AGCTGAGGAATGTCGTTGTG-3′
19
20
5
134 bp
Enhanced green
fluorescent protein
coat protein of
cucumber fruit
mottle mosaic
virus
235 bp