用酵母双杂交系统筛选与痢疾杆菌侵袭素IpaC相互作用的蛋白  

Identification of a Protein Interacting with Shigella flexneri IpaC Invasin by Yeast Two-hybrid System

作　　者：阎晓宇[1] 姚潇[2] 王恒樑[2] 冯尔玲[2] 史兆兴[2] 苏国富[2] 游松[1] 黄留玉[2] 

机构地区：[1]沈阳药科大学制药工程学院,沈阳110016 [2]北京生物工程研究所,北京100071

出　　处：《Acta Genetica Sinica》2004年第4期369-374,共6页

基　　金：首都二四八重大创新工程 (编号 :H0 10 2 10 3 60 119);国家重点基础研究规划 (973项目;编号 :G19990 5 410 3 )资助~~

摘　　要：利用酵母双杂交技术筛选与IpaC相互作用的真核细胞蛋白。首先将IpaC全长基因克隆到诱饵蛋白载体pGBKT7中 ,以构建的pGBKT IpaC为靶蛋白质粒 ,筛选人HeLa细胞cDNA文库。在 2× 10 6个克隆中得到 2 2个阳性克隆。其中 2个阳性克隆编码人胶原酶蛋白片段。并且进一步证明与胶原酶片段相互作用的IpaC结构域在 6 2aa～ 170aa之间。提示IpaC可能在胶原酶参与的生物学过程中发挥作用。Two-hybrid system was applied to screen proteins interacting with IpaC in the host cell.By using two-hybrid system,the bait plasmid containing ipaC gene was constructed and designated pGBKT-IpaC.A human HeLa cDNA library was screened to isolate protein factors that might interact with IpaC.Among the 2×10~6 clones screened,22 positive clones were picked out.Sequence analysis revealed that two of them contained cDNA fragments from collagenase.Subsequently the domain of IpaC interacting with collagenase fragment was identified.These results suggest that IpaC might play a role in some biological processes where collagenase is involved.

关 键 词：痢疾杆菌 酵母双杂交系统 IPAC 胶原酶 

分 类 号：Q933[生物学—微生物学]