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, Sc hmitt strain) has been determined. We used long RT-PCR to generate full len gth DNA amplicon of CVB6, and then directly sequenced the amplicons. One-st ep cloning of the full length amplicon enabled us to obtain an infectious c lone of CVB6. RNA generated from CVB6 amplicon DNA or CVB6 clones, by trans cription with T7 RNA polymerase, was demonstrated to be infectious upon tra nsfection into HeLa cells in vitro. The CVB6 genome is characteristic of en teroviruses, 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 swi ne vesicular disease virus (SVDV). (C) 1999 Published by Elsevier Science B .V. All rights reserved.