Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins.

Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins.
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DOI:
10.1016/j.jviromet.2009.06.017
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发表时间:
2009-11
影响因子:
3.1
通讯作者:
Matsuda Z
Matsuda Z
中科院分区:
医学4区
文献类型:
--
作者:
Wang J;Kondo N;Long Y;Iwamoto A;Matsuda Z

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一个简单的,基于细胞的,膜融合检测系统,使用分裂绿色荧光蛋白(spGFPs)作为指示剂的开发。将pleckstrin同源(PH)结构域连接到每个spGFP的N-末端不仅将报告信号定位到质膜,而且还有助于17个氨基酸残基的较小spGFP的稳定表达。结果表明,该系统允许实时监测膜融合的HIV-1包膜蛋白(Env),而无需添加外部底物。该方法也适用于其他病毒膜融合的分析。
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.
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