文献类型：专利

英文题名：CAMP genetically encoded fluorescent probe useful in CAMP Detection; for monitoring changes in intracellular cyclic live nucleotides, comprises fluorescent protein sequence or its derivatives and cAMP-sensitive polypeptide

作者：XU L;YANG Y;ZHANG Q

机构：[1]UNIV EAST CHINA SCI & TECHNOLOGY

申请号：CN105646716-A

申请日：2014-11-14

公开日：2016-06-08

语种：英文

收录：DERWENT

摘要：NOVELTY - CAMP genetically encoded fluorescent probe comprises fluorescent protein sequence or its derivatives and cAMP-sensitive polypeptide, where the cAMP-sensitive polypeptide is (1) CNBD domain having cAMP binding characteristic, and/or (2) derived from cAMP to cAMP-sensitive is activated directly regulated guanine nucleotide exchange factor Epac family of proteins. USE - The fluorescent probe is useful in CAMP Detection; for monitoring changes in intracellular cyclic live nucleotides (all claimed). ADVANTAGE - The fluorescent probe has excellent specificity, sensitivity and intuition. DETAILED DESCRIPTION - INDEPENDENT CLAIMS are also included for: (1) gene encoding fluorescent probe nucleotide sequence comprising encoding cAMP-sensitive protein nucleotide sequence and encoding fluorescent protein nucleotide sequence; (2) preparing a fluorescent probe comprising (i) adding expression vector containing nucleotide sequence into a host cell, (ii) culturing the host cells under conditions for expression of the host cell, and (iii) separating the host cells from the fluorescent probe; (3) fusion protein comprising fluorescent probe, where the fusion protein is fluorescent probe and specific subcellular localization signals are fused together, the positioning signal may be positioned within the target protein specific organelles; (4) encoding fusion protein nucleotide sequence; (5) preparing expression vector comprising operatively connecting vector plasmid pRSETb and nucleic acid sequence; (6) use of fluorescent probe in high throughput screening processing comprising in pRSETb empty blank control bacteria, cpYFP as a negative control, cAMP expression of a small amount of the fluorescent probe is titrating after closing bacteria with Hepes buffer, resuspending and diluting to same concentration, sonicating, then diluting supernatant to specified concentration detection experiments; and (7) reagent kit comprising specification and fluorescent probe.