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. 2012 Apr;86(7):3541–3553. doi: 10.1128/JVI.06992-11

Fig 5.

Fig 5

Construction and replication of ORF3 and ORF75 deletion viruses. (A) Representation of the BACmid Bac43MOD encompassing the HVS strain C488 genome. The locations of the mini-F plasmid, the EGFP expression cassette, and orf3 and orf75 are indicated. (B) Schematic details of the deletions introduced into the recombinant viruses. (C) Pulsed-field gel electrophoresis of XhoI-digested BACmids. (D) Expression of viral early ORF50/RTA, late ORF17/protease, and ORF3 and ORF75 in infected OMK cells detected by immunoblotting using specific rabbit polyclonal antibodies and swine antirabbit horseradish peroxidase conjugate. (E) Delayed lytic replication of BACmid-derived HVS with orf3 and orf75 deletions compared to replication of the wild-type virus. Viral genome copy numbers in the supernatant of infected cells were quantified by a quantitative PCR detecting the viral orf25 gene at the indicated time points after infection of OMK cells at a low MOI (MOI, 0.05).