Fig. 5.

Mutational analysis for revertant chimeric viruses and vTLV8. (A) Semi-quantitative RT-PCR analysis of vTLV8 and revertants. Total cellular RNAs were extracted from mutant plasmids-transfected BHK-21 cells at 24 h post transfection. β-Actin was used as an internal control and pAS as an unreplicative control. (B) Semi-quantitative analysis of the relative abundance of the negative-strand genomic RNAs and sg mRNAs in transfected BHK-21 cells. The RT-PCR products presented in (A) were quantified by using the Quantity One 1-D analysis software version 4.6.2. The transcription level of the parental vAPRRS was set at 100% and the mock value was subtracted. (C) Indirect immunofluorescence assay. The infected MARC-145 cell monolayers were fixed at 24 h post infection and stained with monoclonal antibody against N protein.