柞蚕核型多角体病毒ptp-2基因克隆及表达  

Cloning and expression of Antheraea pernyi nucleopolyhedrovirus ptp-2 gene

作　　者：石生林[1,2] 潘敏慧[1] 鲁成[1] 

机构地区：[1]西南大学农业部蚕桑学重点实验室 [2]沈阳农业大学生物科学技术学院,辽宁沈阳110161

出　　处：《浙江大学学报（农业与生命科学版）》2007年第2期125-128,共4页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家重点基础研究发展计划973"资助项目(2005CB121000);国家科技攻关计划资助项目(2005BA711A07).

摘　　要：为研究柞蚕核型多角体病毒(Antheraea pernyi nucleopolyhedrovirus,ApNPV)ptp-2基因特性,根据ApNPV PstⅠ-C片段核酸序列,设计引物克隆了ApNPV ptp-2基因;将ptp-2基因亚克隆至原核表达载体pQE-30,构建重组质粒pQE-ptp-2,并在大肠杆菌M15中进行诱导表达,表达量达菌体总蛋白的38.7%,经His标签杂交鉴定证明表达产物为His融合蛋白.以原核表达的His-PTP-2蛋白作为抗原,制作兔多克隆抗血清,Western blot分析表明PTP-2蛋白不参与ApNPV包涵体的结构组成.To elucidate Antheraea pernyi nucleopolyhedrovirus （ApNPV） ptp-2 gene characters, ApNPV ptp- 2 gene was cloned using primers based on the sequence of ApNPV PstI-C fragment and then sub-cloned into prokaryotic expression vector pQE-30 to construct recombinant plasrnid pQE-ptp-2. PTP-2 protein was induced expression in E. coli M15. The expression accounted for 38. 7 percent of the total protein. Western blot analysis of His-tag indicated that the protein expressed was His-fused. PTP-2 polyclonal antiserum was made against His-PTP-2 protein expressed in Escherichia coli. Western blot analysis of ApNPV occlusion body （OB） revealed that PTP-2 protein was not associated with the structure of ApNPV OB.

关 键 词：核型多角体病毒 基因表达 基因克隆 柞蚕 ptp-2基因 

分 类 号：Q93[生物学—微生物学]