NOSTRIN基因克隆及其在人脐静脉内皮细胞中的表达

目的 :构建内皮型一氧化氮合酶运输介导物 (endothelialnitricoxidesynthasetrafficinducer ,NOSTRIN)基因的真核表达载体 ,通过质粒转染人脐静脉内皮细胞 (humanumbilicalveinendothelialcell,HUVEC)获得高表达NOSTRIN基因的细胞克隆。方法 :构建真核表达载体pcDNA3.1 NOSTRIN ,采用脂质体介导将重组质粒导入体外培养的HUVEC ,Westernblot检测转染细胞和未转染细胞中NOSTRIN蛋白质的表达。结果 :成功构建了真核表达载体pcDNA3.1 NOSTRIN ,并用脂质体介导的方法获得了高稳定表达NOSTRIN的细胞克隆 ;Westernblot显示转染前后的细胞均有 5 8kD的蛋白质表达 ,转染后表达量明显增高。结论 :重组质粒pcDNA3.1 NOSTRIN经转染能在HUVEC细胞中高效表达 ,为进一步研究NOSTRIN对HUVEC的生物学影响奠定了基础

Clone and expression of NOSTRIN gene in human umbilical vein endothelial cell

Objective:To construct eukaryotic expression plasmid of endothelial nitric oxide synthase traffic inducer (NOSTRIN) and establish human umbilical vein endothelial cell (HUVEC) line monoclonal cells with the stable expression of NOSTRIN gene by transfection.Methods:The recombinant expression plasmid pcDNA3.1 NOSTRIN was constructed from pRSET NOSTRIN,then was introduced into HUVEC cell line by liposome method.Western blotting was applied to detect NOSTRIN expression in transfected and non transfected cells.Results:The recombinant expression plasmid pcDNA3.1 NOSTRIN was constructed successfully. In HUVEC cell line transfected by the plasmid. Western blotting confirmed that both cell lines express 58kD NOSTRIN but the level was much higher in transfected cells.Conclusion:NOSTRIN can be expressed high effectively in HUVEC cell lines transfected by recombinant plasmid pcDNA3.1 NOSTRIN. The success of this research provides us with the basis for further study on the NOSTRIN function in HUVEC.