利用酵母双杂交筛选与DAT1的LIM2相互作用的蛋白质  

Identification of DAT1/LIM2-interacting proteins by yeast two-hybrid screening

作　　者：惠玲[1] 季朝能[2] 蔡文琴[1] 顾少华[2] 应康[2] 谢毅[2] 毛裕民[2] 

机构地区：[1]第三军医大学基础医学部组织胚胎学教研室,重庆市神经科学研究所重庆400038 [2]复旦大学生命科学院遗传所,上海200433

出　　处：《第三军医大学学报》2005年第5期410-413,共4页Journal of Third Military Medical University

基　　金：国家自然科学青年基金资助项目(30400125)~~

摘　　要：目的　利用酵母双杂交系统筛选人胎脑组织中能与多巴胺相关转录因子DAT1的LIM2相互作用的蛋白质, 以初步了解DAT1的功能。方法　构建酵母双杂交诱饵质粒pLexA-LIM2,筛选人胎脑cDNA文库,并用酵母交配试验初步 验证。结果　获得3个能与DAT1的第2个LIM结构域LIM2结合的相互作用蛋白,通过酵母结合试验证实了它们在酵母 中的结合。结论　DAT1的第2个LIM结构域LIM2可与唐氏综合征关键区基因2(SimilartoDownsyndromecriticalregion gene2,DSCR2)、钙 整合素结合蛋白(calciumandintegrinbindingprotein,CIB)、和HSPC170(CD34+hematopoieticstem/progenitor cells,HSPCs)结合。Objective To understand the functions of a novel gene, dopamine-associated transcription factor (DAT1). Methods The bait plasmid pLexA-LIM2 was constructed and used to screen the human fetal brain cDNA expressing library using yeast two-hybrid. The interaction between LIM2 and positive clones were verified by yeast mating. Results Three proteins were isolated as interacting proteins of DAT1/LIM2. Yeast mating proved that LIM2 of DAT1 could interact with the three proteins in yeast. Conclusion Calcium and integrin binding protein (CIB), similar to Down syndrome critical region gene 2 (DSCR2) and HSPC170 can interact with DAT1/LIM2 in yeast.

关 键 词：蛋白质 杂交筛选 syndrome INTEGRIN 酵母双杂交系统 protein 相关转录因子 cDNA文库 相互作用蛋白 人胎脑组织 唐氏综合征 cells 诱饵质粒 交配试验 Down 结合蛋白 结构域 LIM 多巴胺 整合素 and 初步 验证 

分 类 号：R151.3[医药卫生—营养与食品卫生学] R735.7[医药卫生—公共卫生与预防医学]