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A plasmid construct is developed with the sequence for GFP2 and DsRed2 fluorescent proteins, linked by a 12-amino-acid-long cleavage recognition site for the 2A protease (2Apro), encoded by the EV71 genome and specific for the members of Picornaviridae family. In the construct expressed in HeLa cells, the linker binds the fluorophores within the F\u00f6rster distance and creates a condition for FRET to occur, thus resulting in shortening of the GFP2 fluorescence lifetime. On cells infection with EV71, viral 2Apro released to the cytoplasm cleaves the recognition site, causing disruption of FRET through separation of the fluorophores. Thus, increased GFP2 lifetime to the native values, manifested by the time-correlated single-photon counting, serves as an efficient and specific indicator of the EV71 virus infection.<\/jats:p>","DOI":"10.1117\/1.2718582","type":"journal-article","created":{"date-parts":[[2007,4,30]],"date-time":"2007-04-30T18:26:10Z","timestamp":1177957570000},"page":"1-8","source":"Crossref","is-referenced-by-count":19,"title":["Application of fluorescence resonance energy transfer resolved by fluorescence lifetime imaging microscopy for the detection of enterovirus 71 infection in cells"],"prefix":"10.1117","volume":"12","author":[{"given":"Vladimir","family":"Gukassyan","sequence":"first","affiliation":[{"name":"National Yang-Ming Univ."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueh-Ying","family":"Hsu","sequence":"additional","affiliation":[{"name":"National Yang-Ming Univ."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Szu-Hao","family":"Kung","sequence":"additional","affiliation":[{"name":"National Yang-Ming Univ."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fu-Jen","family":"Kao","sequence":"additional","affiliation":[{"name":"National Yang-Ming Univ."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"189","reference":[{"key":"10.1117\/1.2718582_r1","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1093\/infdis\/129.3.304","volume":"129","author":"Schmidt","year":"1974","journal-title":"J. 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