贝氏柯克斯体外膜蛋白H(OmpH)原核表达载体的构建与鉴定

Construction and Identification of Outer Membrance Protein H (OmpH) of Coxiella burnetii

本试验旨在构建贝氏柯克斯体外膜蛋白H(outer membrance protein H,OmpH)的重组表达载体并分析该蛋白的免疫原性,以贝氏柯克斯体九里株为模板,通过PCR方法扩增出含798bp的贝氏柯克斯体的OmpH基因片段,并将其克隆至原核表达载体pQE-30,得到重组表达质粒pQE-30/OmpH。经IPTG诱导后,SDS-PAGE结果发现,该重组蛋白大小约为25ku。Western blotting试验结果显示,该方法所诱导产生的重组蛋白具有良好反应原性。

This study was amied to construct and identify the prokaryotic expression plasmids of outer membrance protein H （OmpH） of Coxiella burnetii and analysis its immunogenicity. Amplified OmpH gene from the Nine Miles by PCR, the results showed that the OmpH included 798 hp. Cloned OrnpH gene into prokaryotic expression vector pQE-30 after identified by PCR,enzyme digestion and sequence. Constructed the recombinant plasmid pQE-30/OmpH,and then the plasmid was trans- formed into BL21（DE3） competent cells and induced by IPTG. The results showed that pQE-30/OmpH recombinant proteins were 25 ku by SDS-PAGE, and Western blotting assay indicated that the recombinant protein could express OmpH protein with significant antigencity.