摘要
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
Objective Expressing human CD59 on the membrane of cultured pig aortic endothelial cells (PAECs) by gene transfection technieque.Methods The PAECs were cultured in vitro.The pcDNA3-ICAM-2-Enhancer-CD59cDNA and pcDNA3-ICAM-2-CD59cDNA and pcDNA3-CMV-CD59cDNA expression vector were transfected into the PAECs by lipofectinnmine mediated method.Normal PAECs served as negative control and the human erythrocytes as positive control.Stable transfecttants were selected using G418 (concentration 100 mg/L).The expression of transfected cells was assayed by flow cytometry.Results The expression intensity of CD59 expression was 61.4% in PAECs pool that was transfected by pcDNA3-ICAM-2-Enhancer-CD59cDNA,1.7% in blank control PAECs and 8.2% in fibroblast respectively.The intensity of CD59 expression was 38.3% in PAECs pool that was transfected by pcDNA3-ICAM-2-CD59cDNA.Conclusion Enhancer plus promoter of ICAM-2 could tissue-specially induce recombinant CD59 expression in PAECs.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2004年第10期1192-1193,共2页
Chinese Journal of Experimental Surgery
基金
天津市自然科学基金资助项目 (0 1 361 1 0 1 1 )
