Invitro Replication And Assembly Of Vesicular Stomatitis-Virus Nucleocapsids

An in vitro system using vesicular stomatitis virus (VSV) infected HeLa cell S10 extracts, optimized for transcription and translation, replicated and assembled VSV genome-size RNA into nucleocapsids containing at least N protein. The in vitro synthesized nucleocapsids (RNPs) were RNase resistant and had the same buoyant density in CsCl as virion nucleocapsids. Full-length (+) and (−) RNA were synthesized in about the same ratio as in vivo . In vitro replication was dependent on protein synthesis or at least a pool of VSV RNP proteins. This system will be useful for identification of the host cell factors necessary for replication and the relationship of L and NS proteins in transcription and replication.