PET_(15b)-BCG HSP65-GP_2重组质粒的构建与鉴定  

Construction of recombinant plasmid expressed PET_(15b)-BCG HSP65-GP_2 fusion protein gene

作　　者：王丽娟[1] 吴士彬[2] 于永利[3] 王丽颖[4] 

机构地区：[1]蚌埠医学院病原生物学教研室,安徽蚌埠233003 [2]安徽省蚌埠市第三人民医院泌尿外科,233000 [3]吉林大学基础医学院免疫学教研室,吉林长春130021 [4]吉林大学基础医学院分子生物学教研室,吉林长春130021

出　　处：《蚌埠医学院学报》2002年第5期383-384,共2页Journal of Bengbu Medical College

摘　　要：目的 :构建表达卡介苗热休克蛋白 6 5 (BCGHSP6 5 )与人HER 2 /neu细胞毒T淋巴细胞 (CTL)表位 (GP2 )构成的融合蛋白 (BCGHSP6 5 GP2 )的重组质粒。方法 :将用PCR方法扩增并回收的BCGHSP6 5 GP2 基因片段与原核表达质粒PET15b重组 ,经过酶谱分析筛选出正向重组质粒 ,DNA序列分析正向重组质粒。结果 :筛选出 3个正向重组质粒。结论 :重组表达质粒PET15b BCGHSP6 5 GP2 的构建是成功的 。Objective:To construct a recombinant plasmid that expressed BCG HSP65 fused with human HER 2/neu cytotoxic T lymphocyte(CTL) epitope.Methods:BCG HSP65 GP 2 gene fragment was amplified by PCR approach and then cloned into prokaryote expressed plasmid PET 15b .Forward recombinant plasmids were screened by endo nucleases digestion analysis.Results:Three forward recombinant plasmids were obtained.Conclusions:It has been suggested that we have constructed a recombinant expressed PET 15b vector which expressed BCG HSP65 fused with human HER 2/neu CTL epitope.It lay foundation on further expression and biofunction assay.

关 键 词：PET15b-BCG HSP65-GP2 重组质粒 构建 鉴定 基因表达调节 癌基因产物 热休克蛋白类 HER-2/NEU 

分 类 号：Q343.1[生物学—遗传学] Q782