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    • 3. 发明公开
    • 군소 투-하이브리드 시스템
    • APLYSIA双混合系统
    • KR1020040031277A
    • 2004-04-13
    • KR1020020060660
    • 2002-10-04
    • 재단법인서울대학교산학협력재단
    • 강봉균최정환
    • C12N15/63
    • C12N15/85C12Q1/34G01N33/5058G01N33/6812G01N2333/938
    • PURPOSE: An aplysia two-hybrid system is provided, thereby verifying correlation between transcription factors during memory process, so that it can be useful for the examination of mind disease causing factors and screening of drugs relating to correlation between proteins. CONSTITUTION: An aplysia two-hybrid system comprises (a) a reporter vector 4xGal4-LacZ containing Gal4 binding domain and LacZ gene; (b) pNEX-Gal4DB-bait vector containing Gal4 DNA binding domain-bait polynucleotide in pNEX vector; and (c) pNEXδ-Gal4AD prey vector containing Gal4 transcription activation domain in pNEXδ vector, wherein the bait and prey polynucleotides encode transcription factors during memory process; the bait and prey polynucleotide is selected from ApCREB1a, ApCREB2 and ApC/EBP. A method for examining correlation between proteins comprises micro-injecting the aplysia two-hybrid system into an aplysia nerve cell, and analyzing beta-galactosidase from the aplysia nerve cell through X-gal dyeing.
    • 目的:提供aplysia双杂交系统,从而验证记忆过程中转录因子之间的相关性,从而可用于检查心脑疾病引起因素和筛选与蛋白质之间的相关性相关的药物。 构成:aplysia双杂交体系包含(a)含有Gal4结合结构域和LacZ基因的报道载体4xGal4-LacZ; (b)pNEX载体中含有Gal4 DNA结合域诱饵多核苷酸的pNEX-Gal4DB-诱饵载体; 和(c)pNEXδ载体中含有Gal4转录激活结构域的pNEXδ-Gal4AD猎物载体,其中诱饵和猎物多核苷酸在记忆过程中编码转录因子; 诱饵和猎物多核苷酸选自ApCREB1a,ApCREB2和ApC / EBP。 一种检测蛋白质相关性的方法包括将aplysia双杂交系统微注射到神经细胞,并通过X-gal染色分析来自神经细胞膜神经细胞的β-半乳糖苷酶。