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[目的]构建人轮状病毒糖蛋白VP7基因毕赤酵母表达系统。[方法]构建好的毕赤酵母重组表达质粒VP7-pPICZαA经SacⅠ线性化后应用电转化法转化毕赤酵母菌株GS115感受态中,Zeocin平板筛选,PCR鉴定转化子,MDH和MMH平板鉴定Mut表型。[结果]线性化的重组质粒VP7-pPICZα成功转化进入毕赤酵母感受态中,PCR鉴定结果与预期相符,表型确定为Mut+。[结论]成功构建人轮状病毒糖蛋白VP7基因的毕赤酵母表达系统
[Objective] To construct the Pichia expression system of human rotavirus glycoprotein VP7 gene. [Method] The recombinant Pichia pastoris recombinant plasmid VP7-pPICZαA was transformed into Pichia pastoris strain GS115 by electroporation after linearization by SacⅠ. The Zeocin plate was screened by electroporation, and transformants were identified by PCR, MD and MMH plates were identified by Mut type. [Result] The linearized recombinant plasmid VP7-pPICZα was successfully transformed into Pichia pastoris. The result of PCR was consistent with the expectation. The phenotype was identified as Mut +. [Conclusion] The Pichia expression system of human rotavirus glycoprotein VP7 gene was successfully constructed