The complete consensus sequence of coxsackievirus B6 and generation of infectious clones by long RT-PCR.
The full length sequence for the human pathogen coxsackievirus B6 (CVB6, Schmitt strain) has been determined. We used long RT-PCR to generate full length DNA amplicon of CVB6, and then directly sequenced the amplicons. One-step cloning of the full length amplicon enabled us to obtain an infectious clone of CVB6. RNA generated from CVB6 amplicon DNA or CVB6 clones, by transcription with T7 RNA polymerase, was demonstrated to be infectious upon transfection into HeLa cells in vitro. The CVB6 genome is characteristic of enteroviruses, with a 5'-non-translated region (743 nucleotides) followed by an open reading frame (encoding a 2184 amino acid polyprotein) and a 3'-non-translated region (100 nucleotides) and polyadenylated tail. The predicted amino acid sequence of CVB6 clustered with the other CVB serotypes and swine vesicular disease virus (SVDV).
Martino
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TA
Center for Cardiovascular Research, Toronto Hospital, 200 Elizabeth Street, EC12-324, Toronto, Ont., Canada.
Tellier
R
R
Petric
M
M
Irwin
D M
DM
Afshar
A
A
Liu
P P
PP
D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D016384:Consensus Sequence; D017931:DNA Primers; D004770:Enterovirus / Q000145:classification / Q000235:genetics; D029822:Enterovirus B, Human / Q000145:classification / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D017931:DNA Primers; D012367:RNA, Viral