Reassembled GFP: detecting protein-protein interactions and protein expression patterns.
Reassembled GFP: detecting protein-protein interactions and protein expression patterns.
复制标题
重组 GFP：检测蛋白质-蛋白质相互作用和蛋白质表达模式。
DOI：
10.1002/0471739499.ch17
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Regan,Lynne
中科院分区：
文献类型：
--
作者：
Magliery,ThomasJ;Regan,Lynne
Genomic research has resulted in the identification of tens of thousands of putative proteins from all three domains of life in recent years, many of which have no clear function. Key clues to the function of these proteins come from identifying their binding partners and expression patterns. Therefore, it is now of critical importance to develop robust, highthroughput methods to address these issues (Zhu et al., 2003). Immunoprecipitation and related methods like TAP-TAG (Puig et al., 2001) require purification of the analyte protein, demand relatively strong interactions between protein partners, and are not amenable to library approaches. Fusions to Aequorea victoria GFP and its variants have been used to examine expression patterns (Chalfie et al., 1994) and protein interactions through fluorescence resonance energy transfer (FRET)(Miyawaki et al., 1997), but these methods are limited by the photophysical properties of GFP variants and the promoters available to drive expression, particularly in whole organisms. Several combinatorial screens based on the reassembly of dissected proteins have been introduced in recent years to determine the identity of protein ligands, beginning with the yeast two-hybrid screen. In the last five years, experiments have demonstrated that GFP and its variants can be dissected and reassembled to yield fluorescent products. GFP reassembly can be used to demonstrate and identify protein–protein interactions and protein expression patterns in cells and whole organisms.
