Efficient bunyavirus rescue from cloned cDNA

被引：87

作者：

Lowen, AC ^{[1
]}

Noonan, C ^{[1
]}

McLees, A ^{[1
]}

Elliott, RM ^{[1
]}

机构：

[1] Univ Glasgow, Div Virol, Inst Biomed & Life Sci, Glasgow G11 5JR, Lanark, Scotland

来源：

VIROLOGY | 2004年 / 330卷 / 02期

基金：

英国惠康基金;

关键词：

bunyavirus; cDNA; Bunyaviridae; virus rescue; reverse genetics;

D O I：

10.1016/j.virol.2004.10.009

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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, 1540015404) 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 >10(7) 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. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：493 / 500

页数：8

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