桑泛素基因的克隆和表达  

Cloning,Sequence Analysis and Prokaryotic Expression of the Ubiquitin Gene of Morus bombycis

作　　者：高维锡[1,2] 庄艳[1] 常润磊[3] 邹宁[2,4] 

机构地区：[1]烟台职业学院,山东烟台264670 [2]山东师范大学生命科学学院,山东济南250014 [3]华南农业大学林学院,广东广州510642 [4]鲁东大学生命科学学院,山东烟台264025

出　　处：《烟台大学学报（自然科学与工程版）》2009年第4期264-267,共4页Journal of Yantai University(Natural Science and Engineering Edition)

摘　　要：根据桑(Morus bombycis)泛素基因编码区的序列(DQ839402.)设计特异引物,用RT-PCR方法克隆桑泛素基因的编码区.序列分析表明,该编码区长240 bp,编码79个氨基酸,软件分析推测编码蛋白的相对分子量和等电点分别为8.74 kD和7.16.通过同源建模获得了桑泛素基因编码蛋白的理论三维结构.将桑泛素基因与pET-32a(+)连接,构建原核表达载体pET-32a-ub,经IPTG诱导,在大肠杆菌BL21(DE3)中获得高效表达,并经western bolt印迹证明实现了原核表达,为进一步研究其作用机理奠定基础.The coding sequence of Morus bombycis ubiquitin gene is cloned and determined.The opening reading frame（ORF） is 240 bp in length,encoding 79 amino acids with the molecular weight of 8.74 kD and theoretical isoelectric point of 7.16.The theoretical three-dimensional structure of this gene is displayed by homology modeling.Using pET-32a（＋） as a fused expression vector,a recombinant plasmid pET-32a-ub containing the coding sequence of M.bombycis ubiquitin gene is constructed.Western blot indicates that the M. bombycis ubiquitin gene is expressed successfully in the BL21 （DE3） strain of E. coli induced with IPTG. The results may serve as basis for further study on the function of ubiquitin gene in M. bombycis.

关 键 词：桑 泛素 基因克隆 基因序列分析 原核表达 

分 类 号：Q78[生物学—分子生物学]