Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
11029055 | Virology | 2018 | 11 Pages |
Abstract
Recombinant SHFV infectious cDNA clones expressing a foreign gene from an additional sg mRNA were constructed. Two 3â² genomic region sites, between ORF4â² and ORF2b and between ORF4 and ORF5, were utilized for insertion of the myxoma M013 gene with a C-terminal V5 tag followed by one of the three inserted transcription regulatory sequences (TRS), TRS2â², TRS4â² or TRS7. M013 insertion at the ORF4â²/ORF2b site but not the ORF4/ORF5 site generated progeny virus but only the recombinant virus with an inserted TRS2â² retained the entire M013 gene through passage four. Insertion of an auto-fluorescent protein gene, iLOV, with an inserted TRS2â² at the ORF4â²/ORF2b site, generated viable progeny virus. iLOV expression was maintained through passage eight. Although regulation of SHFV subgenomic RNA synthesis is complex, the ORF4â²/ORF2b site, which is located between the two sets of minor structural proteins, is able to tolerate foreign gene insertion.
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Authors
Han Di, Esther K. Morantz, Heena Sadhwani, Joseph C. Jr., Margo A. Brinton,