细粒棘球绦虫肌动蛋白基因的克隆表达  

Cloning and expressing the actin gene of Echinococcus granulosus

作　　者：高瑞雪 李超群[1] 张耀刚 李凤辉[1] 赵婧[1] 姜博璠 张发斌[1] GAO Ruixue;LI Chaoqun;ZHANG Yaogang;LI Fenghui;ZHAO Jing;JIANG Bofan;ZHANG Fabin(Qinghai University Medical College,Xining,Qinghai 810001;Qinghai Provincial Key Laboratory for Echinococcosis,Xining,Qinghai 810001)

机构地区：[1]青海大学医学院,青海西宁810001 [2]青海省包虫病研究重点实验室,青海西宁810001

出　　处：《中国高原医学与生物学杂志》2020年第2期111-114,124,共5页Journal of Chinese High Altitude Medicine & Biology

基　　金：青海大学医学院中青年科技项目(2017-QYY-5)。

摘　　要：目的克隆表达细粒棘球绦虫肌动蛋白(EgACT)基因,并获得EgACT纯化蛋白.方法将经全序列合成、密码子优化合成的目的基因EgACT克隆入PET28a-SUMO表达载体中、转化入Rosetta(DE3)中,诱导表达采用异丙基-β-D-硫代半乳糖苷(IPTG),经镍琼脂糖凝胶亲和层析法获得纯化蛋白.结果重组质粒PET28a-SUMO-EgACT构建成功.SDS-PAGE结果显示,目的蛋白在Rosetta(DE3)中获得高效表达,相对分子量约为60 kDa.结论完成细粒棘球绦虫肌动蛋白基因的克隆及其在Rosetta(DE3)中的表达,并获得纯化蛋白.Objective To clone and express the Echinococcus granulosus actin(EgACT)gene and obtain EgACT purified protein.Methods Through complete sequence synthesis and codon optimization,the target gene EgACT was synthesized.It was cloned into PET28a-SUMO expression vector,transformed into Rosetta(DE3),and induced to express by isopropyl-β-D-thiogalactoside(IPTG).With the method of Nickel Agarose gel afnity chromatography to obtain purified protein.Results The recombinant plasmid PET28a-SUMO-EgACT was sucessfully constructed.The SDS-PAGE resuls showed that the target protein was highly expressed in Rosetta(DE3)with a relative molecular weight 60 kDa approximately.Conclusion We are scesfully cloned the Echinococcus granulosus actin gene and expressed it in Rosetta(DE3).

关 键 词：细粒棘球绦虫 肌动蛋白 基因 克隆 原核表达 

分 类 号：R383.3+3[医药卫生—医学寄生虫学]