Efficient bunyavirus rescue from cloned cDNA

Lowen, A.C., Noonan, C., McLees, A. and Elliott, R.M. (2004) Efficient bunyavirus rescue from cloned cDNA. Virology, 330, pp. 493-500. (doi: 10.1016/j.virol.2004.10.009)

Abstract

Bunyaviruses are trisegmented, negative-sense RNA viruses. Previously, we described a rescue system to recover infectious Bunyamwera virus (genus Orthobunyavirus) entirely from cloned cDNA (Bridgen, A. and Elliott, R.M. (1996) Proc. Nat. Acad. Sci. USA 93, 15400–15404) utilizing a recombinant vaccinia virus expressing bacteriophage T7 RNA polymerase to drive intracellular transcription of transfected T7 promoter-containing plasmids. Here we report efforts to improve the efficiency of the system by comparing different methods of providing T7 polymerase. We found that a BHK-derived cell line BSR-T7/5 that constitutively expresses T7 RNA polymerase supported efficient and reproducible recovery of Bunyamwera virus, routinely generating >107 pfu per rescue experiment. Furthermore, we show that the virus can be recovered from transfecting cells with just three plasmids that express full-length antigenome viral RNAs, greatly simplifying the procedure. We suggest that this procedure should be applicable to viruses in other genera of the family Bunyaviridae and perhaps also to arenaviruses.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Elliott, Professor Richard
• Authors: Lowen, A.C., Noonan, C., McLees, A., and Elliott, R.M.
• College/School: College of Medical Veterinary and Life Sciences
• Journal Name: Virology