Fig. 3

Engineering CPSMV RNA2 cDNA into a vector for VIGS and nonviral gene expression. A Diagram of the RNA2 cDNA under control of the 2X35S promoter, with the amino acid sequences flanking the protease processing site between MP and L-CP shown immediately below (10 aa residues before the processing site, and 20 after). The processing site is denoted with a pair of dark blue scissors. The bottom row shows the FZ constructs with the protease processing site duplicated, and two Eco72I sites introduced for future insertion of GOI fragments. Note that the two protease processing sites are now denoted with light blue scissors, and the duplicated aa residues in gray letters, both reflecting the fact that the nt sequence encoding the duplicated aa residues has been codon-shuffled to minimize nt sequence identity, hence the chance of recombination. B A table listing one control construct (FZ-NC), and two VIGS constructs (FZ-NbPDS and FZ-GmPDS), along with their insert sizes. C Photobleaching in N. benthamiana due to FZ-NbPDS-induced VIGS of NbPDS. D Verification of reduced NbPDS mRNA levels with sqRT-PCR. RT-: control reaction without reverse transcriptase. E Photobleaching in soybean (accession Williams 82) due to FZ-GmPDS-induced VIGS of GmPDS. F Verification of reduced GmPDS mRNA levels with sqRT-PCR