FIG. 1.

Reconstitution of influenza virus mini-RNPs. Cultures of COS-1 cells were infected with vaccinia T7 virus and transfected with pGPB1, pGPB2, pGPA, and pGNPpolyA plasmids. The viral template was provided by simultaneous transfection of pT7NSΔCAT-RT plasmid or by delayed transfection of vNSZ RNA, as indicated in Materials and Methods. After incubation for 48 h, the viral RNPs were extracted and used for in vitro RNA synthesis by using ApG as primer. The RNA product was isolated and analyzed by electrophoresis on sequencing gels. The product obtained after transfection of pT7NSΔCAT-RT (313 nt) is shown (COMP), as well as the product generated by transfection of vNSZ RNA (240 nt) and those obtained when each of the elements of the systems was omitted. The panel to the left shows a 20× overexposure of the lane containing the vNSZ RNA product. The lengths of molecular weight markers are indicated to the left in nucleotides.