人内皮细胞特异性分子-1的转染及其在ECV304中的表达

Transfection of endothelial cell-specific molecular-1 and its expression in ECV304 cells

目的 构建人内皮细胞特异性分子 1(ESM 1)真核表达载体 ,转染人脐静脉内皮细胞系ECV30 4 ,并在ECV30 4中获得表达。方法 将ESM 1cDNA插入真核载体pIRES的多克隆位点中 ,转染人脐静脉内皮细胞。原位杂交 ,免疫组化检测ESM 1基因的表达。结果 构建的表达载体经限制性内切酶酶切图谱分析和DNA序列分析证实 ,转染后发现ESM 1的表达在转录和蛋白水平都明显的增加。结论 成功构建了人ESM 1真核表达载体和转染了人脐静脉内皮细胞系ECV30 4 ,并获得高水平的表达 ,为进一步研究作好准备。

Objective To construct endothelial cell-specific molecular-1(ESM-1) eukaryotic express vector,transfect it into human umbilical vein endothelial cells( HUVECs,ECV304) and express it in vitro . Methods ESM-1 cDNA was inserted into eukaryotic vector pIRES and transfected into HUVECs. The ESM-1 expression was detected by in situ hybridization,immunohistochemistry.Results The recombinant plasmid was identified by restriction endonucleases mapping analysis and DNA sequencing. The expression of ESM-1 was obviously increased in the level of transcription and translation.Conclusion The eukaryotic express vector is constructed and transfected into HUVECs successfully. High level of expression of ESM-1 is obtained in ECV304. This study lays a good basis for further research.