[目的]构建截短型AMA1的原核表达载体,并进行体外诱导表达,为在体外表达弓形虫AMA1重组蛋白.[方法]设计去掉弓形虫AMA1信号肽的PCR引物,以弓形虫cDNA为模板进行PCR扩增,PCR扩增产物酶切后与pGEX-4T-3原核表达载体连接,并转化到BL21感受态细胞内.对经PCR鉴定为阳性的重组质粒pGEX-4T-3-AMA1进行体外诱导表达.[结果]成功构建了弓形虫AMA1的原核表达质粒pGEX-4T-3-AMA1,通过体外诱导表明,AMA1融合蛋白是以包涵体的形式存在.[结论]弓形虫AMA1蛋白的体外表达为进一步制备抗AMA1血清及免疫学实验奠定了基础.%[Objective] The aim was to construct recombinant prokaryotic expression vector of truncated AMA1 fragment and induced expression in vitro was carried out. [Methods] The PCR was performed using T. Gondii cDNA as templates, and then the production of PCR was connected with pGEX-4T-3 prokaryotic expression vector. The recombinant vector with AMA1 was transferred into BL21 competent cells, and was introduced in vitro. [Result] We successfully constructed recombinant prokaryotic expression vector pGEX-4T-3-AMAl which could express the AMA1 fusion protein in vitro. It confirmed that the AMA1 fusion protein was the inclusion body exist in E. Coli. [Conclusion] These results provided foundation for the further studies on the immunologic tests for T. Gondii.
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