基于瞬时转染的病毒蛋白融合检测法
Transient Transfection-based Fusion Assay for Viral Proteins.
作者信息
Vallbracht Melina, Schröter Christina, Klupp Barbara G, Mettenleiter Thomas C
机构信息
Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
出版信息
Bio Protoc. 2017 Mar 5;7(5):e2162. doi: 10.21769/BioProtoc.2162.
Abstract
Membrane fusion is vital for entry of enveloped viruses into host cells as well as for direct viral cell-to-cell spread. To understand the fusion mechanism in more detail, we use an infection free system whereby fusion can be induced by a minimal set of the alphaherpesvirus pseudorabies virus (PrV) glycoproteins gB, gH and gL. Here, we describe an optimized protocol of a transient transfection based fusion assay to quantify cell-cell fusion induced by the PrV glycoproteins.
摘要
膜融合对于包膜病毒进入宿主细胞以及病毒在细胞间的直接传播至关重要。为了更详细地了解融合机制，我们使用了一种无感染系统，通过该系统，可由一组最少的甲型疱疹病毒伪狂犬病病毒（PrV）糖蛋白gB、gH和gL诱导融合。在此，我们描述了一种基于瞬时转染的融合测定优化方案，用于量化由PrV糖蛋白诱导的细胞间融合。
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本文引用的文献
1
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J Virol. 2015 Dec 9;90(5):2264-72. doi: 10.1128/JVI.02739-15.
2
Viral membrane fusion.
Virology. 2015 May;479-480:498-507. doi: 10.1016/j.virol.2015.03.043. Epub 2015 Apr 10.
3
Analysis of fusion using a virus-free cell fusion assay.
Methods Mol Biol. 2005;292:187-96. doi: 10.1385/1-59259-848-x:187.
4
Characterization of cell-cell fusion mediated by herpes simplex virus 2 glycoproteins gB, gD, gH and gL in transfected cells.
J Gen Virol. 2000 Aug;81(Pt 8):2017-2027. doi: 10.1099/0022-1317-81-8-2017.
5
Glycoproteins gB, gD, and gHgL of herpes simplex virus type 1 are necessary and sufficient to mediate membrane fusion in a Cos cell transfection system.
J Virol. 1998 Jan;72(1):873-5. doi: 10.1128/JVI.72.1.873-875.1998.
6
Adaptability in herpesviruses: glycoprotein D-independent infectivity of pseudorabies virus.
J Virol. 1997 Jan;71(1):17-24. doi: 10.1128/JVI.71.1.17-24.1997.
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