稳定表达饰胶蛋白聚糖基因的大鼠肾小球系膜细胞株的建立  

Constitution of Rat Glomerular Mesangial Cell Stably Expressing Decorin Gene

作　　者：王慧君[1] 张志刚[1] 陈广平 林文生[1] 陈琦[1] 郭慕依[1] 

机构地区：[1]复旦大学基础医学院病理学教研室,上海200032 [2]美国北海岸LIJ健康机构放射肿瘤学部,纽约11030

出　　处：《复旦学报（医学版）》2001年第2期97-99,102,共4页Fudan University Journal of Medical Sciences

基　　金：上海市教委重点学科基金!(B990 80 2 )资助项目

摘　　要：目的　研究将饰胶蛋白聚糖 (decorin ,DCN)基因转染大鼠肾小球系膜细胞的可行性。方法　RT PCR法扩增大鼠DCN基因 ,构建重组真核表达质粒 pcDNA3.1A DCN ,采用脂质体将其转入大鼠MsC ,经G418筛选阳性克隆 ,Westernblot及RT PCR法鉴定。结果　成功构建重组真核表达质粒 pcDNA3 .1A DCN ,转染MsC并筛选出 2个阳性克隆株。结论　构建载有DCN基因的MsC载体 。Purpose: To study the possibility of transferring decorin gene to rat glomerular mesangial cell. Methods: Amplification of the rat decorin (DCN) cDNA by RT-PCR for constructing the plasmid pcDNA3. 1A-DCN and lipofectin method for transfecting DCN gene into MsC; G418 scanning, Western blot and RT-PCR analysis for detecting DCN protein and mRNA in D-A6 cell clone. Results: The recombinant eukaryotic expression plasmid, pcDNA3. 1A-DCN was successfully constructed and 2 cell clones positively expressing DCN were selected. Conclusions: These cell clones positively expressing DCN is valuable for providing favorable experimental bases of gene therapy to model with glomerular diseases.

关 键 词：肾小球系膜细胞 饰胶蛋白聚糖 基因转染 大鼠 DCN 

分 类 号：R692.602[医药卫生—泌尿科学]