豚鼠衣原体GPIC0474多克隆抗体制备

Preparation of Polyclonal Antibodies against GPIC0474 of Chlamydia Caviae

目的对GPIC0474进行克隆、重组、表达并制备GPIC0474的多克隆抗体,鉴定GPIC0474是否与其相似基因肺炎衣原体包涵体膜蛋白CPn0308具有相似的定位。方法通过PCR扩增GPIC0474,构建pGEX-6P2-GPIC0474重组质粒并转化,筛选阳性克隆提取质粒进行测序分析。对重组质粒进行诱导表达,以GST-GPIC0474为免疫原,对Balb/C小鼠进行常规免疫,分离血清获得多克隆抗体。结果成功制备了GPIC0474的多克隆抗体。结论试验所制备的GPIC0474的多克隆抗体为研究GPIC0474的定位提供了材料,进而为以后研究GPIC0474的功能奠定了基础。

Objective To identify whether GPIC0474 has the similar position with that of Chlamydia pneumoniae inclusion membrane protein CPn0308. In the experiment, GPIC0474 gene was cloned, re- combined and expressed. GST fusion protein was prepared. Then polyclonal antibodies （pAbs） to GST- GPIC0474. were made. Methods The target gene GPIC0474 was amplified by PCR, and then recombi- nant plasmid pGEX-6P2-GPIC0474 was constructed and transferred. The positive colonies were selected and plasmid was sequenced and analyzed. Then the recombinant was induced and GST fusion protein was expressed. The purified fusion protein was used to immunize mice in a routine way, pAbs to the fusion protein was obtained by separating serum from whole blood. Results preparation of polyclonal antibodies against GPIC0474 was successful. Conclusion pAbs to GPIC0474 fusion protein provides material for lacation of GPIC0474. Thus it lays foundation for studying the function of GPIC0474.