人轮状病毒外衣壳糖蛋白VP7基因毕赤酵母重组表达系统的构建  

CONSTRUCTION OF VP7 GENE OF HUMAN ROTAVIRUS IN PICHIA PASTORIS EXPRES-SION SYSTEM

作　　者：卢颖[1] 佟伟[1] 单颖[1] 张轶博[1] 吴囡[1] 董颖[1] 李华[1] 吴学敏[1] 

机构地区：[1]辽宁医学院免疫与病原生物学教研室,锦州121001

出　　处：《现代预防医学》2010年第18期3523-3524,3528,共3页Modern Preventive Medicine

摘　　要：[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+。[Objective] To construct VP7 gene of Human Rotavirus glycoprotein in Pichia pastoris expression system.[Methods] Recombination expression plasmid VP7-pPICZαA was transformed to the competent Pichia pastoris GS115 cells by using electroporation method after being linear by SacⅠ.The recombinant transformants were screened with Zeocin and identified by PCR.Phenotype was identified by using MDH and MMH plate.[Results] The linear recombinant plasmid pPICZαA-VP7 were transformed into competent Pichia pastoris successfully.The recombinant strains were proven correct after identification with PCR,and their phenotypes were Mut＋.[Conclusion] The expression system of human rotavirus VP7 gene in Pichia pas-toris was successfully constructed.

关 键 词：轮状病毒 VP7基因 毕赤酵母 

分 类 号：R373[医药卫生—病原生物学]