Figure 10. Analysis of the splicing efficiency of the truncated forms of the ORF1 and ORF2 mutants by RT-PCR.

(A) Schematic representation of a JCV Mad-1 late transcript illustrating the insertion of a stop codon (TAA) at position after T524 (GTT⁵²⁴ TAA C⁵²⁵ATGG) to block the expression of the unique amino acid regions of both ORF1 and ORF2 proteins. (B) RT-PCR analysis of the JCV late transcripts isolated from SVG-A cells either untransfected (− Cont.) or transfected/infected with JCV Mad-1 WT or Mad-1 T524 stop mutant using the following primers: 5′-primer (1469-1489) and 3′-primer (780-755) as described in Materials and Methods. The total RNA was also subjected to RT-PCR for amplification of GAPDH RNA as described for Fig. 6A. Amplified products were then analyzed on a 3% agarose gel.