过表达ERCC1蛋白细胞系的建立  

Establishment of over-expressing ERCC1 protein cell line

作　　者：陈新志[1] 洪新[1] 李海山[1] 卢永科[1] 杜伯雨[1] 仲来福[1] 

机构地区：[1]大连医科大学毒理学研究室,辽宁大连116027

出　　处：《大连医科大学学报》2003年第3期163-168,共6页Journal of Dalian Medical University

摘　　要：[目的 ]建立过表达ERCC1蛋白的细胞系 ,为遗传毒性化合物的研究提供模型细胞系 ,并有可能为具有DNA加合作用化合物提供新的毒理学筛选方法。 [方法 ]应用PCR和DNA重组技术将核苷酸切除修复交叉互补组 1 (Excisionrepaircrosscomplementationgroup1 ,ERCC1 )基因与pT7Blue载体相连接 ,然后构建真核细胞重组表达质粒 pcDNA3.1 /Zeo(+) -ERCC1 ,并导入 2 93细胞。经Zeocin抗性筛选后获得转染ERCC1基因的细胞克隆。细胞增殖后以Western -blot检测目的蛋白的表达。 [结果 ]获得稳定转染ERCC1基因的细胞系 ,并在细胞中检测出有ERCC1蛋白的过表达。 [结论 ]用基因转染法成功建立了过表达ERCC1蛋白的ERCC1 - 2To establish an over-expressing ERCC1 protein model cell line ERCC1-293 to be used for study of genotoxic chemicals. Excision repair cross complementation group1(ERCC1) gene was transferred to pT7Blue vector with polymerase chain reaction (PCR) and DNA recombinant techniques. Recombinant express plasmid pcDNA3.1/Zeo(+)-ERCC1 was constructed and introduced into 293 cells. The ERCC1 transfected cell clones were obtained with Zeocin selection. The expression of ERCC1 protein was detected by Western-blot after the cells being proliferated. The model cell line transfected with ERCC1 was gained, which could over-express the ERCC1 protein in cells. [Conclusion] An over-expressing ERCC1 protein model cell line is established successfully by gene transfection method.

关 键 词：ERCCL 蛋白细胞系 基因表达 转染 遗传毒性化合物 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]