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目的 :克隆并表达含有刚地弓形虫主要表面抗原P30及霍乱毒素A2 /B亚基基因的原核表达载体 ,为弓形虫疫苗的研究奠定基础。方法 :通过PCR方法扩增出P30基因片段 ,将其克隆入含有霍乱毒素A2 /B亚基基因的表达质粒pUAB0 2 4 ,在大肠杆菌JM10 9(DE3)中表达融合蛋白。行SDS PAGE电泳及West ernblotting检测鉴定。结果 :酶切电泳证明质粒构建正确。SDS PAGE显示IPTG诱导可以产生特异性条带。Westernblotting进一步证实该条带为p30 CTA2 /B融合蛋白。结论 :成功构建的表达载体pUAB0 2 4 p30可有效表达特异性的融合抗原蛋白P30 CTA2 /B。
OBJECTIVE: To clone and express the prokaryotic expression vector containing the major surface antigens of Toxoplasma gondii P30 and cholera toxin A2 / B subunit gene, and lay the foundation for the study of Toxoplasma gondii vaccine. Methods: The P30 gene fragment was amplified by PCR and cloned into the expression plasmid pUAB024 containing the cholera toxin A2 / B subunit gene. The fusion protein was expressed in E. coli JM109 (DE3). SDS PAGE electrophoresis and West ernblotting detection. Results: Electrophoresis showed that the plasmid was constructed correctly. SDS PAGE showed that IPTG induction can produce specific bands. Westernblotting further confirmed that the band was p30 CTA2 / B fusion protein. CONCLUSION: The successfully constructed expression vector pUAB0 2 4 p30 can effectively express the specific fusion antigen protein P30 CTA2 / B.