贝氏柯克斯体精氨酸受体基因的原核表达及免疫原性分析

Prokaryotic Expression and Immunogenicity Analysis of Arginine Repressor Gene of Coxiella burnetii

构建贝氏柯克斯体精氨酸受体(argR)的重组表达载体并分析该重组蛋白的免疫原性。应用PCR方法,以贝氏柯克斯体九里株为模板,扩增出含734bp的精氨酸受体目的基因片段,并将其克隆至原核表达载体pGEX-6p-1,得到重组表达质粒pGEX-6p/argR,经IPTG诱导表达后产生ArgR重组蛋白,SDSPAGE分析该蛋白大小约为25ku,Western blot结果显示,表达的重组蛋白有着良好的免疫原性。用argR免疫小鼠,能诱发特异性抗体的产生,说明该原核表达蛋白具有良好的免疫原性。

This study is aimed to construct and identify the prokaryotic expression plasmids of arginine re-pressor gene（argR）of Coxiella burnetii and analyze its immunogenicity.The argR from Nine Mile strain was amplified by PCR,argR fragment is 734 bp.The argR gene was cloned into the prokaryotic expression vector pGEX-6p-1 after identification by PCR,enzyme digestion and sequencing.The recombinant plasmid pGEX-6p/argR was constructed,and then the plasmid was transformed into BL21（DE3）competent cells and induced by IPTG.The result showed that pGEX-6p/argR recombinant protein is 25 ku.SDS-PAGE and Western blot indicated that the recombinant protein could express argR protein with significant antigenci-ty.The protein argR induced specific antibodies in immunized mice and had good immunoreactivity.