重组土拉弗朗西斯菌外膜蛋白FopA的融合表达及抗原性分析

Expression and Antigenicity Analysis of Recombinant Francisella Outer Membrane Protein FopA

目的:探索一种大量表达功能性土拉弗朗西斯菌外膜蛋白FopA的方法。方法:采用SignalP 3.0 Server进行信号肽预测,将土拉弗朗西斯菌外膜蛋白FopA信号肽的基因序列(75bp)去除,将1107bp的核心序列克隆至原核表达载体pET32a,并在大肠杆菌BL21(DE3)中诱导表达。结果:构建了pET32a-fopA载体,重组蛋白FopA表达量约占菌体总蛋白量60%,Western blot分析显示重组FopA蛋白有较好的抗原性。结论:获得了高效表达FopA的pET32a-fopA表达载体,为下一步土拉弗朗西斯菌外膜蛋白FopA应用研究奠定了基础。

Objective：To search for a new method to achieve greater levels of expression and production of large quantities of functional Francisella outer membrane protein FopA.Methods：According to the result of SignalP 3.0 Server,the native leader sequence of gene fopA was removed by PCR and genetic recombinant technique.The DNA fragment（1107bp）encoding the functional FopA（without its signal sequence）was cloned into pET32a and expressed in E.coli BL21（DE3）.Results：The prokaryotic expression vector pET32a-fopA was constructed and the level of recombinant FopA induced by IPTG was up to 60%in expression.Western blot showed that recombinant FopA was antigenic.Conclusion：The efficient vector of pET32a-fopA was produced,which set good basis for the further applied study of recombinant FopA.