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J Virol Methods. 2009 Nov;161(2):216-22. doi: 10.1016/j.jviromet.2009.06.017. Epub 2009 Jun 25.

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

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China-Japan Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Chaoyang District, Beijing, China.


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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