鼠疫耶尔森菌外膜蛋白的克隆与表达

Cloning and expression of outer membrane proteins of Yersinia pestis

摘要:
目的克隆并表达鼠疫耶尔森菌的多种外膜蛋白或外膜蛋白的定位、表达调控蛋白。
方法通过PCR技术对目的基因进行扩增, 其PCR产物经纯化、酶切处理后, 再分别导入原核表达载体pET32a中, 然后转化大肠埃希菌BL21(DE3), IPTG诱导后, 进行聚丙烯酰胺凝胶电泳(SDS-PAGE)分析和蛋白质芯片分析。
结果获得了23个表达鼠疫耶尔森菌外膜蛋白或外膜蛋白的定位、表达调控蛋白的克隆子.全部重组蛋白质的纯度均在85%以上。
结论构建了一系列重组表达质粒, 为进一步研究鼠疫耶尔森菌外膜蛋白的结构与功能奠定了基础。

Abstract:
Objective To clone and express the outer membrane proteins of Yersinia pestis.
Methods The genes encoded the outer membrane proteins of Yersinis pestis were amplified by PCR and PCR products were digested by restriction enzymes.Then the restricted fragments were ligated to the prokaryotic expression vector p ET32a and transformed into E.coli.After inducing with 0.1mM IPT G, SDS-PA GE and protein chip were used to detect proteins that were expressed.
Results All of the selected genes were expressed in E.coli.with a level of 20-45% of the total bacterial proteins.The purity of expressed products were up to 85%.
Conclusion This work settled a foundation for studying the structure and function of the outer membrane proteins Yersinia pestis.