使用修饰的分裂绿色荧光蛋白监测HIV-1包膜介导的膜融合
Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins.
作者信息
Wang Jianqi, Kondo Naoyuki, Long Yufei, Iwamoto Aikichi, Matsuda Zene
机构信息
China-Japan Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Chaoyang District, Beijing, China.
出版信息
J Virol Methods. 2009 Nov;161(2):216-22. doi: 10.1016/j.jviromet.2009.06.017. Epub 2009 Jun 25.
Abstract
A simple, cell-based, membrane fusion assay system that uses split green fluorescent proteins (spGFPs) as an indicator was developed. The attachment of the pleckstrin homology (PH) domain to the N-termini of each spGFP not only localized the reporter signal to the plasma membrane but also helped the stable expression of the smaller spGFP of seventeen amino acid residues. It was shown that this system allowed real-time monitoring of membrane fusion by HIV-1 envelope protein (Env) without the addition of external substrates. This method can be adapted to the analyses of other viral membrane fusion.
摘要
开发了一种简单的基于细胞的膜融合检测系统，该系统使用分裂绿色荧光蛋白（spGFP）作为指示剂。将普列克底物蛋白同源（PH）结构域连接到每个spGFP的N端，不仅将报告信号定位到质膜，还有助于17个氨基酸残基的较小spGFP的稳定表达。结果表明，该系统无需添加外部底物即可实时监测HIV-1包膜蛋白（Env）介导的膜融合。该方法可适用于其他病毒膜融合的分析。
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