用酵母双杂交检测BAR与PDZ结构域的相互作用  

Interaction between BAR and PDZ Domain Assayed by Yeast Two-hybrid System

作　　者：肖虹[1] 袁静明[1] 石亚伟[1] 

机构地区：[1]山西大学生物技术研究所化学生物学与分子工程教育部重点实验室,山西太原030006

出　　处：《山西大学学报（自然科学版）》2006年第4期421-424,共4页Journal of Shanxi University(Natural Science Edition)

基　　金：国家自然科学基金(30400065);山西省留学基金项目

摘　　要：用酵母双杂交系统探索P ICK 1分子中BAR结构域与PDZ结构域的相互作用.分别构建了pGAD-PDZ和pGBK-BAR重组子,共转入酵母细胞后,涂布于SD(L eu-,T rp-,H is-)固体平板培养基,经滤纸复印和显色反应,在8 h内观察到只有pGAD-PDZ/pGBK-BAR的表达产物呈阳性,而其它对照pGADT 7/pGBKT 7、pGAD-PDZ/pGBKT 7和pGADT 7/pGBK-BAR均呈阴性反应,说明BAR与PDZ结构域具有相互作用.The interaction between BAR domain and PDZ domain in PICK1 were studied by using yeast two-hybrid system. Two recombinant plasmids, pGAD-pdz and pGBK-bar were constructed and in turn, co-introduced into yeast AH109. Subsequently, SD （Leu^-, Trp^-,His^-）solid plate medium was used to grow and screen transformed yeast. The colony were copied with whatman No. 5 filter paper and coloured with Z-buffer/X-gal. The result showed that the expresed product of the recombine yeast containing pGAD-PDZ/pGBK-BAR rerealed the position reaction as compared other controls pGADT7/pGBKT7, pGAD-PDZ/pGBKT7 and pGADT7/pGBK-BAR. It suggested that the interaction between BAR domain and PDZ domain was occurred.

关 键 词：PICK1 BAR结构域 PDZ结构域 酵母双杂交系统 相互作用 

分 类 号：Q814[生物学—生物工程]