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

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,Western blot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,为其开展对肾小球疾病模型的基因治疗提供了良好的实验基础。

Constitution of RatGlomerular Mesangial Cell Stably Expressing Decorin Gene

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.