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目的 探讨外源性A2 0基因在内皮细胞获得稳定表达的可行性。 方法 将N 末端标记E tag的HA2 0cDNA重组于pCAGGS真核表达载体 ,构建了pCAGGSEHA2 0真核表达重组体。DOTAP脂质体介导的pCAGGSEHA2 0和pMAMneo基因转染 ,经G4 18筛选阳性克隆 ,再用免疫荧光及分子原位杂交检测A2 0基因的表达。 结果 成功构建了pCAGGSEHA2 0真核表达重组体 ,A2 0基因在经G4 18筛选后的内皮细胞中得到有效表达。 结论 在DOTAP脂质体介导下 ,A2 0基因能够导入内皮细胞并在其内获得表达
Objective To investigate the feasibility of stable expression of exogenous A2 0 gene in endothelial cells. Methods The HA2 0 cDNA of N-terminal tagged E tag was recombined with pCAGGS eukaryotic expression vector and a recombinant plasmid pCAGGSEHA20 was constructed. DOTAP liposome-mediated transfection of pCAGGSEHA20 and pMAMneo genes, positive clones were screened by G418, and the expression of A2 0 gene was detected by immunofluorescence and molecular in situ hybridization. Results The eukaryotic expression recombinant pCAGGSEHA20 was successfully constructed. The A2 0 gene was effectively expressed in endothelial cells screened by G418. Conclusion The A2 0 gene can be introduced into endothelial cells and expressed in DOTAP liposomes