摘要
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 construct Prokaryotic expression plasmid containing major surface antigen P30 of toxoplasma gondii and cholera toxin A 2/B subunit gene to express fusion protein. Methods: P30 gene. Fragment was Obtained by PCR and inserted into express plasmid PUAB024 which contains cholera toxin A2/B subunit gene. Fusion protein was expressed in JM109(DE3) and SDS-PAGE and Western blot were adopted for detection and appraisal. Results: Correct construction of express plasmid was proved by SDS-PAGE,which showed that specific band could be produced through IPTG induction.The band was further verified by western blotting as fussion: protein P30-CTA2/B. Conclusion: Prokaryotic express plasmid PUAB024 was successfully constructed and the fusion protein P30-CTA2/B was specifically expressed.
出处
《山东大学学报（医学版）》
CAS
2002年第3期210-212,共3页
Journal of Shandong University:Health Sciences
基金
山东省自然科学基金资助项目 (Y99C0 2 )
关键词
弓形虫属
霍乱毒素
基因表达
大肠杆菌
Toxoplasma
Cholera toxin
Gene expression
Escherichia coli
