鹦鹉热嗜衣原体CPSIT-0531融合蛋白原核表达载体的构建、表达及免疫原性检测  

Construction and expression of the prokaryotic clone of recombinant protein CPSIT-0531 of Chlamydophila psittaci and detection of its immunongenicity

作　　者：贺庆芝[1] 曾怀才[1] 陆春雪[1] 王川[1] 吴移谋[1] 

机构地区：[1]南华大学病原生物研究所,湖南衡阳421001

出　　处：《实用预防医学》2014年第2期129-132,共4页Practical Preventive Medicine

基　　金：国家自然科学基金(81202323);湖南省科技厅项目(2012FJ6009);湖南省卫生厅项目(B2012-042)

摘　　要：目的构建鹦鹉热嗜衣原体(Chlamydophila psittaci,Cps)CPSIT-0531原核表达载体,表达并纯化该融合蛋白,检测其免疫原性。方法采用PCR技术从Cps 6BC基因组中扩增CPSIT-0531基因,并构建pET30a-CPSIT-0531原核表达载体,然后转化到宿主菌E.coli BL21中,IPTG诱导His-CPSIT-0531融合蛋白表达,免疫BALB/c小鼠制备多克隆抗体,ELISA分析His-CPSIT-0531蛋白的免疫原性。结果成功构建了pET30a-CPSIT-0531原核表达载体,并表达和纯化较稳定的重组蛋白;GST-CPSIT-0531免疫小鼠后,ELISA检测免疫小鼠的血清特异性抗体效价为1:640 000。结论成功克隆表达了His-CPSIT-0531,该蛋白具有较好的免疫原性。Objective The aim was to construct the prokaryotic expression plasmid of CPSIT-0531 gene of Chlamydophila psittaci （Cps） 6BC strain, exprexs and purify the recombinant protein His - CPSIT-0531 and detect its immunongenicity. Methods CPSIT-0531 gene was cloned by PCR and inserted into the expression vector pET30a to construct pET30a- CPSIT- 0531 recombinant plasmid. The recombinant plasmid was transformed into E. coli BL21 to express His- CPSlT-0531 fusion protein. The protein was purified and used to immunize BALB/c mice. ELISA were carried out to identify the immunogenicity. Results The recombinant expression plasmid pET30a - CPSIT-0531 was successfully constructed. The His - CPSIT-0531 fu- sion protein was successfully expressed and purified. ELISA showed the specific antibody titer against CPSIT-0531 reached 1 ： 640,000. Conclusions CPSIT-0531 protein is successfully expressed in prokaryotic expression system and it has good immu- nongenicity.

关 键 词：鹦鹉热嗜衣原体 CPSIT_0531 克隆表达 免疫原性 

分 类 号：R374.2[医药卫生—病原生物学]