eNOS基因在JAR细胞中的表达  

Expression of transfected eNOS gene in human JAR cell

作　　者：沈政[1] 铁国栋[1] 刘为敏[1] 赵兴绪[2] 段恩奎[1] 

机构地区：[1]中国科学院动物研究所生殖生物学国家重点实验室,北京100080 [2]甘肃农业大学动物医学院,兰州730070

出　　处：《动物学报》2004年第2期258-262,共5页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金(No30170112);中科院知识创新工程和(NoKSCX3IOZ07)生殖生物学国家重点实验室客座研究经费;甘肃省教育厅重点项目(No012d06)资助~~

摘　　要：本研究应用脂质体介导转染技术将人内皮型一氧化氮合酶 (eNOS)基因转入绒癌细胞系JAR细胞 ,获得转染阳性细胞。用RT PCR和Westernblot技术从基因及蛋白水平对表达产物进行鉴定 ,结果显示 :有较高水平的mRNA转录和特异目的蛋白表达 ;通过免疫细胞化学方法证实 ,转染eNOS基因的阳性JAR细胞与对照组相比 ,有外源eNOS蛋白高表达 ;但是一氧化氮合酶 (NOS)活性及其催化产物NO并没有直接升高 ;使用A2 3187处理则能增加NOS的活性 ,说明在转染的JAR细胞中 。Activity of endothelial nitric oxide synthase(eNOS)is regulated in a complex fashion by eNOS “cofactor-accumulating” enzymes as well as more than 20 interacting proteins.The present study was designed to determine the enzyme activity of eNOS in the transfected JAR cell.The results showed that mRNA of eNOS in the transfected JAR cell significantly increased compared to untreated cells.Immunocytochemistry revealed a strong green perinuclear immunofluorescence in eNOS transfected JAR cell.NOS activity and NO release did not change in transfectants.In this study,three independent methods demonstrated transfection resulted in over-expression of eNOS in JAR cell,but the enzyme was not active until treated with A23187.

关 键 词：ENOS基因 JAR细胞 表达 内皮型一氧化氮合酶 酶活性 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]