大肠杆菌脑微血管内皮细胞侵袭蛋白-yijP的表达纯化  

Expression and purification of yijP protein contributing to E.coli invasion in brain microvascular endothelial cells

作　　者：王淳[1] 方文刚[1] 陈誉华[1] 

机构地区：[1]中国医科大学基础医学院发育生物学研究室

出　　处：《中国医科大学学报》2005年第1期16-18,共3页Journal of China Medical University

摘　　要：目的:研究大肠杆菌脑微血管内皮细胞侵袭基因yijP的编码产物在大肠杆菌侵袭脑微血管内皮细胞中的作用,以期阐明大肠杆菌穿过血脑屏障的机制。方法:应用PCR方法获取yijP基因C末端 1. 04kb片断(编码333个氨基酸),将其克隆到PQE载体并转化到受体菌M15,以构建出表达N 末端带有 6个组氨酸的yijP融合蛋白的工程菌。通过Ni NTAAgarose亲和层析法提纯yijP融合蛋白,初步分析该蛋白对人脑微血管内皮细胞 (HB -MEC)生长的影响。结果:成功获得了分子量为 41kd的yijP融合蛋白,并发现yijP蛋白对HBMEC有较强的细胞毒作用。结论:大肠杆菌脑微血管内皮细胞侵袭基因yijP的编码产物 yijP蛋白可能是一种细胞毒因子。Objective: To study the biological function of the yijP gene contributing to Escherichia coli invasion in brain microvascular endothelial cells(HBMEC). Methods: The 1.04 kb DNA fragment encoding yijP C-terminal 333 amino acids was inserted to pQE vector, and the recombinant plasmid encoded yijP fusion protein with his-tag at N-terminal was constructed. The recombinant plasmids were transferred into M15(pREP4) strain for expressing the yijP fusion protein. The yijP fusion protein expressed was purified by using Ni-NTA agarose affinity chromatography. The function of yijP fusion protein was analyzed by observing HBMEC incubated with yijP fusion protein. Results: We obtained the 41kd yijP fusion protein successfully, and found that yijP fusion protein had toxic effect on HBMEC in vitro. Conclusion: The protein encoded by yijP gene contributing to E.coli invasion of brain microvascular endothelial cells may be a cellular toxin.

关 键 词：大肠杆菌 人脑微血管内皮细胞 血脑屏障 yijP基因 

分 类 号：Q786[生物学—分子生物学]