Minigenome RNA expression and L mutants. (A) RT-PCR. Total RNAs were purified from BSR T7/5 cells transfected with pPIV2-GFP, pTM1-P, pTM1-NP, and pTM1-L or mutants. RT-PCRs were carried out as described in Materials and Methods. Ethidium bromide staining of products from the PCRs is shown. Lane 1, untransfected; lane 2, without pTM1-L; lane 3, PIV2 wt; lane 4, C-terminal-50-aa-truncated L (Fig. 3B, CΔ50); lane 5, K2218/2222/G2225A (Fig. 4B, row 4); lane 6, G2225M (Fig. 5A, row 4); lane 7, H1298/R1299A (Fig. 5A, row 7); lane 8, without the RT reaction. (B) Primer extension. The same RNAs were analyzed by primer extension, using a 32P-oligonucleotide representing positions 178 to 160 (Materials and Methods). Lane 1, untransfected; lane 2, without pTM1-L; lane 3, PIV2 wt; lane 4, C-terminal-50-aa-truncated L; lane 5, K2218/2222/G2225A; lane 6, G2225M; lane 7, H1298/R1299A.