低温诱导堆型艾美耳球虫3-1E基因的可溶性表达  

Expression of Soluble Proteins Eimeria.acervulina 3-1E Gene in E.coli under Low Temperature

作　　者：姚鹏[1] 张建军[1] 王黎霞[2] 安健[1] 

机构地区：[1]北京农学院动物科学技术学院,北京102206 [2]北京农业职业学院畜牧兽医系,北京102442

出　　处：《北京农学院学报》2012年第2期39-42,共4页Journal of Beijing University of Agriculture

基　　金：国家自然基金资助项目(31072128)

摘　　要：设计合成特异引物应用RT-PCR技术扩增堆型艾美耳球虫的3-1E基因。通过EcoRⅠ和NotⅠ双酶切,基因连接到表达载体PET28a上,转化入Escherichia coli BL21(DE3)感受态细菌,挑取阳性菌落。经过双酶切和测序鉴定,证明含目的基因的表达载体PET28a构建成功。低温(16℃)诱导表达,经过SDS-PAGE和We-stren Bolt鉴定,表明此蛋白以可溶蛋白在细菌内表达。在诱导50h,IPTG浓度为0.4mmol/L时目的蛋白表达量最大。The sequences of Eimeria. acervulina 3-1E gene were amplified by RT-PCR. The processed 3 1E gene was connected to PET28a,an expression vector by EcoR I and Not I double digestion and transformed into Escherichia coli BL21（DE3）. The recombinant expression vectors was identified by double digestion, and sequenced. The highest soluble expression level of re- combinant protein was obtained after 50h at 16℃ and the concentration of IPTG was 0.4mmol/L.

关 键 词：堆型艾美耳球虫 3-1E 低温诱导 表达 

分 类 号：S852.72[农业科学—基础兽医学]