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.
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~~