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. 2015 Aug 12;10(8):e0135567. doi: 10.1371/journal.pone.0135567

Fig 11. Transfection of glycoprotein mutant KO GPCMV BACs onto GPL cells.

Fig 11

Individual mutant GPCMV BACs were separately transfected onto GPL fibroblast cells to regenerate virus. GFP reporter gene encoded in the viral genome enabled real time tracking of the development of virus from individual transfected cells. Glycoprotein mutant GPCMV BACs were either transfected individually (panels A, C, E, G, I, K and L) or in combination with a rescue plasmid encoding a wild type locus to restore the mutant back to wild type phenotype where GFP virus could be detected spreading across the cell monolayer (panels B, D, F and H). The gH mutant was also transfected onto a cell line expressing gH in trans to support virus growth (panel J). A gH rescue virus was also generated by co-transfection with a rescue locus plasmid (data not shown). Panels K and L show the outcome for a gO knockout mutant based on the back drop of a virus carrying (L) or lacking (K) epithelial cell tropism. Only gO mutant GPCMV with epithelial tropism could grow on GPL cells. A rescue virus of panel K was generated by co-transfection of the gO GPCMV mutant with a wild type locus plasmid to restore wild type virus phenotype (data not shown). Images taken between day 16–18 post transfection.