Cloning and mapping of the potato virus Y genome and its in vitro expression

Abstract

Full-length cDNA of genomic RNA of potato virus Y (PVY) was cloned in one piece into a lambda vector. The order of the EcoRI and SalI fragments of the inserted cDNA was determined. This is the first report of the cloning of a long, expressible, potyvirus genome. The availability of such a clone is a prerequisite for any further study of the molecular biology of this group of viruses, as they are expressed into a self-processed primary polyprotein.

Plants transformed with a cistron of a potato virus Y protease (NIa) are resistant to virus infection.

“…pGNIal.4 was subcloned into the plasmid pBluescript KS (Stratagene) and transcripts directed by a T7 promoter were used as probes in hybridization assays ofplant material. DNA (20) and RNA (21) promoter and the transcription-termination signal of gene 7 of pTiA6 (13). The sequence at the top of each illustration is that of a translation-initiation signal ligated to the PVY clone.…”