作 者: ; (马进); (简清); (李新辉); (罗建仁); (张红军); (林文辉);
机构地区: 中国水产科学研究院珠江水产研究所
出 处: 《中山大学学报论丛》 1998年第6期70-73,共4页
摘 要: 为了研究鱼生长激素基因在大肠杆菌中的表达,采用聚合酶链反应(PCR)技术对鲑鱼生长激素cDNA的5端和3端进行定向修饰.将修饰后的三种基因分别克隆到大肠杆菌表达质粒pBV220,构建成重组鲑鱼生长激素表达质粒pBVGH11,pBVGH18和pBVGH24,转化大肠杆菌DH5α进行诱导表达.结果表明:核糖核蛋白体结合位点(SD)与起始密码子ATG之间的距离对生长激素的表达水平影响极大.鲑鱼生长激素基因的终止密码子TAG不能有效终止该基因在大肠杆菌中翻译的进行,造成部分通读.加入大肠杆菌强终止子TAA可避免通读和产生超长蛋白。 In order to study fish GH cDNA expression in E.coli , we use PCR method to modify salmon GH cDNA 5and 3ends. Three modified genes were cloned to E.coli expression vector pBV220, constructed salmon GH cDNA expression vectors pBVGH11, pBVGH18 and pBVGH24. After transformation and expression, the results of experiment show that the distance between SD sequence and initial codon ATG is very important to fish GH expression level and salmon GH cDNAs stop codon TAG can not stop E.coli translation effectively. Addition of E.coli strong stop codon TAA can avoid expression of super long protein.