A reverse genetics system for dsRNA viruses that generates birnavirus, such as infectious bursal disease virus, from synthetic RNA transcripts

摘要

This system is for the generation of live Birnavirus such as infectious bursal disease virus (IBDV), a segmented double-stranded (ds)RNA virus of the Birnavirdae family, using synthetic transcripts derived from cloned DNA. Independent full-length cDNA clones were constructed which contained the entire coding and non-coding regions of RNA segments A and B of IBDV, respectively. Synthetic RNAs of both segments were produced by in vitro transcription of linearized plasmids with T7 RNA polymerase. Transfection of Vero cells with combined plus-sense transcripts of both segments generated infectious virus as early as 36 hours post-transfection. The development of a reverse genetics system for dsRNA viruses will facilitate studies of the regulation of viral gene expression pathogenesis, and design of a new generation of live and inactivated vaccines.