小鼠B7-DC胞外段原核表达载体的构建及表达  

Construction and expression of prokaryotic vector encoding extracellular region of murine B7-DC

作　　者：樊燕[1] 江文正[1] 张亚丽[1] 闻洁君[1] 郝文丽[1] 钱旻[1] 

机构地区：[1]华东师范大学生命科学学院,上海200062

出　　处：《细胞与分子免疫学杂志》2008年第3期225-227,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：上海市青年科技启明星计划资助项目(06QA14017)

摘　　要：目的:构建小鼠B7-DC胞外段基因的原核表达载体,并在E.coli BL21中诱导表达。方法:从小鼠骨髓来源的未成熟树突状细胞(DC)中提取总RNA,经RT-PCR扩增B7-DC胞外段,并将其克隆至原核表达载体pET32a(+)中,构建重组表达质粒pET32a(+)-B7-DCECD。重组质粒经双酶切鉴定及序列测定后,转化E.coli BL21,经IPTG诱导表达目的蛋白,并用SDS-PAGE和Western blot进行检测。结果:获得全长为582bp的小鼠B7-DC胞外段基因,经测序证实其序列正确。SDS-PAGE和Western blot分析证实重组质粒可表达出Mr约为41000的B7-DC胞外段蛋白。结论:成功地构建了小鼠B7-DC胞外段基因原核表达载体,并在E.coli BL21中进行表达,为进一步研究B7-DC的功能奠定实验基础。AIM:To construct a prokaryotic expression vector for the extracellular domain of murine B7-DC(B7-DCECD)gene,and to express the gene in E.coli BL21.METHODS:The total RNA was extracted from murine immature bone marrow-derived dendritic cells and the extracellular fragment of B7-DC cDNA was amplified by RT-PCR.The recombinant plasmid pET32a(+)-B7-DCECD was constructed by cloning the extracellular fragment of B7-DC cDNA into the prokaryotic expression vector pET32a(+).After the recombinant plasmid was identifie...

关 键 词：B7-DC 基因克隆 原核表达 

分 类 号：R392.11[医药卫生—免疫学]