In vitro synthesis of the full-length complement of the negative-strand genome RNA of vesicular stomatitis virus.

Abstract

Under the normal conditions of in vitro RNA synthesis, the virion-associated RNA polymerase of vesicular stomatitis virus synthesizes 5 monocistronic mRNA and a 48-nucleotide-long leader RNA that represents the exact 3''-terminal region of the genome RNA. When the transcribing core was preincubated with ATP and CTP, reisolated, and then incubated in the presence of the .beta.,.gamma. imido analog of ATP (AdoPP[NH]P) and the 3 normal ribonucleoside triphosphates, the full-length complementary strand of the genome RNA was synthesized in vitro. Specific phosphorylated states of regulatory proteins may control transcription in vitro to generate the full-length plus strands.