重组pEGFP-C2-p100质粒构建及表达  

Construction and expression of a pEGFP-C2-p100 recombinant plasmid

作　　者：何津岩[1] 东莉洁[1] 葛林[1] 赵钢[1] 邵洁[1] 陆燕欣[1] 李晓冬[1] 姚智[1] 杨洁[1] 

机构地区：[1]天津医科大学免疫学教研室,天津300070

出　　处：《天津医科大学学报》2008年第2期135-137,共3页Journal of Tianjin Medical University

基　　金："863"国家高技术研究发展计划(2007AA02Z115);国家教育部新世纪人才支持计划(NCET-04-0245);国家自然科学基金(30670441;30300070);天津市科委应用基础研究重点项目(07JCZD-JC07300);天津市科委国际合作项目(05YFGHHZ01300)资助项目

摘　　要：目的:将人类p100基因定向连入pEGFP-C2质粒,使p100蛋白可与绿色荧光蛋白在HeLa细胞内融合表达,为进一步研究p100蛋白的功能及定位奠定实验基础。方法:采用EcoRI和BamHI双酶切方法,从pSG5-p100质粒中获得p100蛋白的cDNA全长;将该cDNA连接入pEGFP-C2质粒。将构建成功的pEGFP-C2-p100质粒转染入HeLa细胞,荧光显微镜下观察绿色荧光蛋白表达。结果:(1)将该质粒进行双酶切鉴定可见p100片段。(2)转染重组质粒后可观察到绿色荧光蛋白的表达。结论:(1)p100 cDNA全长成功载入pEGFP-C2质粒。(2)p100蛋白可与绿色荧光蛋白在HeLa细胞中融合表达。Objective： To construct eukaryotic green fluorescent protein （GFP） expressing recombinant plasmids, pEGFP-C2-p100, which contain human p100 full length cDNA. Methods： The p100 full length cDNA was purified after pSG5-p100 plasmids were digested by EcoR Ⅰ and BamH Ⅰ. And then they were inserted into pEGFP-C2 fluorescent expressing vector. We detected the p100 fragment by double digestion. This pEGFP-C2-p100 recombinant plasmids was transfected into HeLa cell line and the expression of green fluorescent protein was observed. Results： （ 1 ）The p100 fragment was detected in the products of the restriction enzyme digestion. （2）The green fluorescent protein could be observed by fluorescence microscope after the transfection. Conclusion： （ 1 ）The full length of p100 cDNA fragment is inserted into the pEGFP-C2 vector. （2）The fluorescent expressing recombinant plasmids pEGFP-C2-p100 could express p100-GFP fusion proteins successfully .

关 键 词：人类p100蛋白 PEGFP-C2 重组质粒 融合蛋白 

分 类 号：Q7[生物学—分子生物学]