用含霍乱毒素A2/B的载体在大肠杆菌中表达弓形虫主要表面抗原P30

目的：克隆并表达含有弓形虫P30抗原及霍乱毒素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：应用PCR方法扩增出P30基因片段后，克隆入含有霍乱毒素A2／B亚基基因的表达质粒pUAB024，在大肠杆菌JM109(DE3)中表达融合蛋白。通过SDS-PAGE电泳及Western blotting进行检测鉴定。结果：电泳证明质粒构建正确，SDS-PAGE显示经IPTG诱导可以产生特异性条带，Western blotting进一步证实该条带为P30-CTA2／B融合蛋白。结论：表达载体pUAB024-P30可有效表达特异性的融合弓形虫抗原蛋白P30-CTA2B。

Construction and expression of toxoplasma major surface antigen P30 fusion protein with cholera toxin A2/B in Bacillus coli

Aim To Constructed prokaryotic expressing plasmid pCT-P30 containing Toxoplasma gondii major surface antigen P30 and cholera toxin A 2B subunit gene and express P30-CTA 2/B fusion protein.Methods Obtained P30 gene fragment by PCR and inserted it to plasmid pUAB024 containing cholera toxin A 2/B subunit gene,Expressed fusion protein in JM109(DE3) and detected by running SDS-PAGE and Western blotting.Results Constructed correctly the expressing plasmid.The expression products were assayed by SDS-PAGE,which showed a specific protein brand and were further confirmed by western blotting showing that the specific fusion protein contained P30.Conclusion Prokaryotic expressing plasmid pUAB024-P30 can effectively express fusion protein P30-CTA 2/B.