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Detection of protein interactions based on GFP fragment complementation by fluorescence microscopy and spectrofluorometry
dc.contributor.author | Torrado, Mario | |
dc.contributor.author | Iglesias Reinoso, Raquel | |
dc.contributor.author | Mikhailov, Alexander T. | |
dc.date.accessioned | 2018-05-15T09:44:35Z | |
dc.date.available | 2018-05-15T09:44:35Z | |
dc.date.issued | 2008 | |
dc.identifier.citation | Torrado M, Iglesias R, Mikhailov AT. Detection of protein interactions based on GFP fragment complementation by fluorescence microscopy and spectrofluorometry. Biotechniques. 2008;44(1):70-74 | es_ES |
dc.identifier.issn | 0736-6205 | |
dc.identifier.issn | 1940-9818 | |
dc.identifier.uri | http://hdl.handle.net/2183/20705 | |
dc.description.abstract | [Abstract] We have developed a set of simple modifications of the green fluorescent protein (GFP)fragment reassembly assay in bacteria that permits: (i)fluorescent microscopy visualization of GFP reassembly only 1-2 h after induction of protein expression, thus approximating the detection of GFP reassembly to the real-time dynamics of protein complex formation in living cells; (ii) spectrofluorometric detection of reassembled GFP fluorescent signals directly in lysates from cell suspension thereby avoiding, in many cases, the need for tag-affinity isolation of protein complexes; and (iii) comparative quantification of signal intensity in numerous cell-sample lysates using a Bio-Rad IQ5 spectrofluorometric detection system (Bio-Rad Laboratories, Madrid, Spain). Collectively, the results demonstrate that the combination of microscopic and spectrofluorometric detection provides a time-saving and sensitive alternative to existing methods of fluorescence complementation analysis. | es_ES |
dc.description.sponsorship | Ministerio de Educación y Ciencia; SAF2004-01462 | |
dc.description.sponsorship | Galicia. Consellería de Innovación, Industria e Comercio; PGIDIT04BTF161001PR | |
dc.language.iso | eng | es_ES |
dc.publisher | Future Science | es_ES |
dc.relation.uri | http://dx.doi.org/10.2144/000112685 | es_ES |
dc.subject | Escherichia coli | es_ES |
dc.subject | Genetic complementation test | es_ES |
dc.subject | Green fluorescent proteins | es_ES |
dc.subject | Peptide fragments | es_ES |
dc.subject | Microscopy-fluorescence | es_ES |
dc.subject | Protein binding | es_ES |
dc.subject | Protein interaction mapping | es_ES |
dc.subject | Spectrometry-fluorescence | es_ES |
dc.title | Detection of protein interactions based on GFP fragment complementation by fluorescence microscopy and spectrofluorometry | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.access | info:eu-repo/semantics/openAccess | es_ES |
UDC.journalTitle | Biotechniques | es_ES |
UDC.volume | 44 | es_ES |
UDC.issue | 1 | es_ES |
UDC.startPage | 70 | es_ES |
UDC.endPage | 74 | es_ES |
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