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

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。

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

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.