Cell-free translational analysis of the processing of infectious pancreatic necrosis virus polyprotein

Abstract

A cDNA clone of the large genomic segment of infectious pancreatic necrosis virus (IPNV) was inserted into transcription vectors and used for production of RNA transcripts of various lengths. These RNA transcripts were used to prime the synthesis of virus-specific polypeptides in a rabbit reticulocyte translation system. Full-length transcripts resulted in the synthesis of processed viral proteins pVP2, NS, and VP3. Transcripts which were deleted in the VP2 coding region did not affect the processing of NS or VP3 and similarly, deletions in VP3 did not affect the formation of NS or VP2. However, deletions which extended into the NS coding region resulted in a loss of protease activity and the production of truncated precursor polypeptides. The virus-specific proteolytic activity could not be inhibited by specific antisera and a trans activity for the viral proteases could not be demonstrated.