稳定高表达人ERP57基因A549细胞株的建立及其对CRT表达的影响  

Cloning and Eukaryotic Expression of Human Recombinant ERP57

作　　者：杨建林[1] 韩钰[1] 王艳林[1] 张伟[1] 

机构地区：[1]三峡大学分子生物学研究所,宜昌443002

出　　处：《生物技术通报》2010年第10期198-200,共3页Biotechnology Bulletin

基　　金：国家自然科学基金(30973445);校级青年科学基金(KJ2008A039)

摘　　要：建立稳定高表达人ERP57蛋白的A549细胞株,并观察对CRT表达的影响。采用巢式RT-PCR从人非小细胞肺腺癌A549细胞总RNA中克隆人ERP57 cDNA,构建ERP57真核表达质粒(pcDNA3.1(+)/ERP57)脂质体转染A549细胞。经G418筛选,获得高表达人ERP57蛋白的A549细胞株。通过Western blotting检测细胞中ERP57及CRT蛋白表达情况。成功获得稳定高表达ERP57蛋白的人A549细胞株,CRT表达量无明显改变。成功获得稳定高表达ERP57蛋白的人A549细胞株,证实细胞中高表达ERP57对CRT表达量无明显影响。The research was to clone,express and purify human recombinant ERP57. Human ERP57 cDNA was amplified from total RNA of human lung cancer cell line A549 cells by RT-PCR. Then,PCR product was sub cloned into eukaryotie expression vector pcDNA3.1 （ ＋ ）. The vector was then transfected into A549 cells by using liposome and the single clones were selected with G418. The cell line that is successful in transfection and secretory expression of ERP57 were identified by Western Blotting assay. Eukaryotic expression plasmid of ERP57 was constructed successfully. After transfected into A549 cells,ERP57 could be expressed.

关 键 词：ERP57 钙网蛋白(CRT) 真核表达 A549细胞 

分 类 号：Q78[生物学—分子生物学]