Analysing host cell shut=off caused by Picorna viruses - by comparing translation prods. of viral 2C gene from RNA with and without cap structure, used e.g. to identify inhibitors of viral 2A protease

摘要

Analysis comprises (1) in vitro transcription of a rhinoviral 2C gene (provided with translation stop and start codons) to produce RNA having a cap structure, (2) adding this to a translation mixt. which has been pretreated with a picorna virus 2A protease, (3) subjecting this mixt. to in vitro translation, and (4) comparing the translation products with those generated in a comparative test using RNA wthout the cap structure. Also new are the plasmids pHRV2/5'UTR-2C, phink-2C, phink-2BC, pSVL-2BC, pSVL-2C and pbeta-2C. Pref. the human rhinovirus (HRV)2 2C-gene is transcribed, and HRV2 2A protease is used in step (2). The translation mixt. includes reticulocyte lysate (or a fraction of it). USE/ADVANTAGE - The method is used (1) to detect cellular portions of the 'host cell shut off' (HCSO) generated proteins, (2) to isolate inhibitors of HCSO, partic. of cellular and viral proteins involved in the process, specifically rhinoviral 2A protease.