Fig 1.
Construction and recovery of pVSVIRES. (A) The circular map shows the structure of plasmid pVSV-MC11-eGFP with artificially inserted unique restriction sites. (B) A schematic representation of VSV genome constructs with IRES elements. The short hairpin is shown before the IRES at the 5′ end. M* indicates a matrix gene mutation (M51 deletion). (C) VSV genome constructs with FMDV and HRV IRES elements. The gene junction is shown in the middle. The pVSVFMDV and pVSVHRV plasmids were made by insertion of FMDV IRES and HRV IRES elements, respectively, before the start codon of the M gene (boxed). The hairpin is also shown (Hp). Capital bold letters are VSV P and M stop and start codons, respectively. The SmaI restriction site is shown in italics and underlined. The nucleotides (40 nt) between P and M ORF are shown as dotted lines (P/M UTR), and the last five nucleotides of the P/M gene junction are also shown (TGTTA). (D) The circular map shows the structure of plasmid pCI-eGFP, in which the eGFP ORF was inserted between NheI and NotI restriction sites. It was driven by the CMV promoter. A short hairpin (Hp) was inserted before the eGFP start codon. (E) Fluorescence microscopic images of BHK cells transfected with equal amounts of pCI-eGFP plasmid with or without hairpin.