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.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
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
