Efficient transcription obtained by a new procedure of the tRNA-like structure of turnip yellow mosaic virus by a template-dependent and specific viral RNA polymerase (vol 64, pg 181, 1997)

The RNA-dependent RNA polymerase (RdRp) of turnip yellow mosaic virus (TYMV) was isolated by a simple, new method. An active, template-dependent and specific enzyme was obtained. Although the genomic RNA of TYMV could not be transcribed completely during an invitro RdRp assay, a complete double-stranded product was identified as being of negative polarity by complete digestion with ribonuclease T1. Antibodies directed to part of the N-terminal (Ab140) or C-terminal (Ab66) invitro autocleavage products of the large non-structural polyprotein of TYMV, could both partially inhibit RdRp activity peaks with different protein compositions. Both peak fractions retained high specificity for transcription of TYMV RNA. A protein of approximately 115 kDa was detected by both Ab140 and Ab66.