论文部分内容阅读
目的 通过转基因技术使猪的血管内皮细胞表达人CD5 9蛋白。方法 体外培养猪血管内皮细胞 ,脂质体转染法将pcDNA3 ICAM 2 Enhancer CD5 9cDNA、pcDNA3 ICAM 2 CD5 9cDNA及 pcDNA3 CMV CD5 9cDNA转染猪的血管内皮细胞 ,以未转染的猪血管内皮细胞为阴性对照 ,人的红细胞为阳性对照 ,经G418筛选 (浓度为 10 0mg/L)获得具有抗性的克隆 ,通过流式细胞仪检测转染细胞的表达情况。结果 pcDNA3 ICAM 2 Enhancer CD5 9cDNA重组质粒阳性表达强度为 61.4% ,对照组为 1.7% ,在成纤维细胞中为 8.2 %。不带增强子质粒为 3 8.3 %。结论 以人细胞间黏附因子Ⅱ (ICAM 2 )为启动子并带增强子的CD5 9重组基因在猪的血管内皮细胞中能高效特异表达
Objective To express porcine CD59 protein in porcine vascular endothelial cells by transgenic technology. Methods Porcine vascular endothelial cells were cultured in vitro. The pcDNA3 ICAM 2 Enhancer CD5 9 cDNA, pcDNA3 ICAM 2 CD5 9 cDNA and pcDNA3 CMV CD5 9 cDNA were transfected into porcine vascular endothelial cells by lipofection. The untransfected porcine vascular endothelial cells were Negative control, human red blood cells as a positive control, the G418 screening (concentration of 10mg / L) to obtain resistant clones, transfected cells by flow cytometry expression. Results The positive expression of pcDNA3 ICAM 2 Enhancer CD5 9 cDNA plasmid was 61.4% in the control group, 1.7% in the control group and 8.2% in the fibroblasts. The plasmid without enhancer was 33.3%. Conclusion The recombinant CD59 gene with ICAM 2 as promoter and enhancer can be efficiently and specifically expressed in porcine vascular endothelial cells