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. 2021 Dec 16;17(12):e1010092. doi: 10.1371/journal.ppat.1010092

Fig 1. Construction of recombinant rVSVInd and rVSVNJ with SF, S1, and RBD+E genes of SARS-CoV-2 with and without honeybee msp and VSV Gtc.

Fig 1

Codon-optimized full-length Spike protein gene (SF), S1 subunit gene and the receptor-binding domain (RBD) plus envelope protein genes of SARS-CoV-2 with and without 21 amino acids honeybee melittin signal peptide [(msp) NH2-MKFLVNVALVFMVVYISYIYA-COOH] [24] gene in the purple box, and 49 amino acids VSV G protein transmembrane domain and cytoplasmic tail [(Gtc) NH2-SSIASFFFIIGLIIGLFL VLRVGIYLCIKLKHTKKRQIYTDIEMNRLGK-COOH] gene in the red box were inserted into the G and L gene junction of rVSVInd and rVSVNJ. In addition, 25- nucleotides-long VSV intergenic junctions (5´-CATATGAAAAAAACTAACAGATATC-3´), in the green box, were inserted between genes to provide transcription termination, polyadenylation and the transcription reinitiation sequences. Recombinant viruses were rescued by VSV reverse genetics [20]. pT7: Bacteriophage T7 promoter for DNA-dependent RNA polymerase. N: VSV Nucleocapsid Protein gene. P: VSV Phosphoprotein gene. M: VSV Matrix protein gene. G: VSV Glycoprotein gene. L: VSV Large protein, RNA-dependent RNA polymerase gene. l: Leader region in the 3´-end of the VSV genome. t: Trailer region in the 5´-end of the VSV genome. HDV: Hepatitis delta virus ribozyme encoding sequences. T7δ: Bacteriophage T7 transcriptional terminator sequences. nt: nucleotides. aa: amino acids.