HCV NS5A反式激活基因NS5ATP13在毕氏酵母中的表达研究  

Expression of NS5ATP13 transregulated by HCV NS5A in Pichia pastoris

作　　者：袁菊[1] 成军[1] 洪源[1] 杨延平[1] 陶明亮[1] 靳亚平[2] 韩莉[1] 

机构地区：[1]北京地坛医院传染病研究所,北京100011 [2]西北农林科技大学动物科技学院

出　　处：《中华实验和临床感染病杂志（电子版）》2007年第4期210-212,共3页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家重点基础研究项目(2004CB518908)

摘　　要：目的探讨NS5AATP13能否在毕氏酵母中成功表达。方法用BstXⅠ将pPICZαA-NS5ATP13线性化后,电转化毕氏酵母菌株KM71H,用Zeocin筛选高拷贝数的酵母菌落,在BMGY和BMMY培养基中进行诱导表达,以甲醇诱导,30℃培养4d,表达产物进行SDS-PAGE和Westernblot分析。结果预期分子量大小的NS5ATP13蛋白分泌至培养液。结论NS5ATP13在毕氏酵母中成功表达。Objective To express NS5ATP13 in Pichia pastoris.Methods The constructed plasmid pPICZαA-NS5ATP13 was linarized with Bst XⅠand transformed into Pichia pastoris KM71H,multi-copy recombinants was selected with Zeocin and expressed in BMGY and BMMY media.The Zeocin-resistant clones were induced expression by methanol,which were growing at 30℃ for 4 days.The expressed products were analyzed by SDS-PAGE and Western blot.Results SDS-PAGE and Western blot showed NS5ATP13 protein with expected molecular weight serected in the supernatant.Conclusions NS5ATP13 protein was successfully expressed in Pichia pastoris.

关 键 词：毕氏酵母 分泌表达 HCV NS5A 

分 类 号：R346[医药卫生—基础医学]