Template-dependent, in vitro replication of rotavirus RNA.
A template-dependent, in vitro rotavirus RNA replication system was established. The system initiated and synthesized full-length double-stranded RNAs on rotavirus positive-sense template RNAs. Native rotavirus mRNAs or in vitro transcripts, with bona fide 3' and 5' termini, derived from rotavirus cDNAs functioned as templates. Replicase activity was associated with a subviral particle containing VP1, VP2, and VP3 and was derived from native virions or baculovirus coexpression of rotavirus genes. A cis-acting signal involved in replication was localized within the 26 3'-terminal nucleotides of a reporter template RNA. Various biochemical and biophysical parameters affecting the efficiency of replication were examined to optimize the replication system. A replication system capable of in vitro initiation has not been previously described for Reoviridae.
Chen
D
D
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030.
Zeng
C Q
CQ
Wentz
M J
MJ
Gorziglia
M
M
Estes
M K
MK
Ramig
R F
RF
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D008969:Molecular Sequence Data; D012330:RNA, Double-Stranded / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D012401:Rotavirus / Q000502:physiology*; D014771:Virion / Q000502:physiology; D014779:Virus Replication
D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D012367:RNA, Viral