棉铃虫核型多角体病毒sod基因在大肠杆菌中的表达  

Expression of HaSNPV sod gene in E. coli

作　　者：高国辉[1] 张传溪[1] 

机构地区：[1]浙江大学应用昆虫学研究所病毒分子生物学实验室,浙江杭州310029

出　　处：《中国病毒学》2003年第5期482-485,共4页Virologica Sinica

基　　金：国家自然科学基金(No.30070032);高等学校全国优秀博士学位论文作者专项资金(No.200055)

摘　　要：用PCR方法从棉铃虫(Helicoverpa armigera)单粒包埋型核型多角体病毒(HaSNPV)C1株基因组中扩增sod基因编码区,克隆到pGEM-T-easy vector,测定了核苷酸序列。将基因编码区克隆到原核表达载体pETblue2,构建了含表达质粒pETblue2/HaSNPV SOD,转化大肠杆菌DE3(BL21)进行IPTG诱导表达。SDS-PAGE分析表明SOD的表达量约为细胞总蛋白的37％。邻苯三酚法测定表达蛋白活性,结果表明每毫克菌体可溶性总蛋白中表达产物校正酶活力单位为694U/mg。The coding region of superoxide dismutase was amplified by using PCR method from Helicoverpa armigera single enveloped nucleocapsid nucleopolyhedrovirus (HaSNPV) C1 genome. The PCR product was cloned into pGEM-T-easy vector and sequenced. The cloned coding region of HaSNPV SOD was inserted into the expression vector pETblue-2 to form the recombinant plasmid pETblue2/HaSNPV SOD and was then transformed into E. coli DE3 for expression. The SDS-PAGE analysis revealed that the expressed recombinant SOD accumulated up to 37% of the total bacterial protein. The enzyme activity of HaSNPV SOD was assayed by Pyrogallol autooxidation method. The result showed that the revised enzyme activity of the expressed product in the total soluble protein was 694 U/mg.

关 键 词：棉铃虫 核型多角体病毒 SOD基因 大肠杆菌 表达载体 

分 类 号：Q786[生物学—分子生物学]