应用二维电泳和质谱技术初步研究NAG7基因的功能(英文)  

Preliminary Function Study of NAG7 Using Two dimensional Electrophoresis and Mass Spectrometry

作　　者：谭琛[1] 李江[1] 谢奕[1] 向秋[1] 王洁如[1] 梁宋平[2] 李桂源[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所,长沙410078 [2]湖南师范大学生命科学学院,长沙410081

出　　处：《生物化学与生物物理学报》2001年第4期373-378,共6页

基　　金：国家 8 6 3高技术研究和发展计划资助(No .10 2 10 0 1 0 5,No .Z19 0 10 1 0 3);国家重点基础研究发展计划资助 (973)(No .G19980 5

摘　　要：为了进一步研究鼻咽癌相关基因NAG7的功能 ,NAG7基因编码框的cDNA片段被亚克隆至 pcDNA3 .1(+ )的表达载体 ,通过脂质体转染入鼻咽癌细胞系HNE1。抽提HNE1细胞和已转染了NAG7基因的HNE1细胞总蛋白质 ,用二维凝胶电泳分离蛋白质 ,获得二维凝胶图谱 ,PDQuest软件包分析后 ,确立表达上调的蛋白质斑点进行质谱分析 ,得到的肽质指纹经蛋白质数据库分析鉴定蛋白质。共分析了 12个蛋白质点 ,其中 3个未有质谱结果 ,已鉴定的 9个蛋白质包括生长捕获特异蛋白、DNA结合蛋白、c myc启动子结合蛋白及胱冬肽酶 (caspase) 6等蛋白质。通过对这些蛋白质性质和功能的讨论 ,发现它们参与了细胞周期调控、转录调节及细胞凋亡等重要事件。因此 。In search of mechanisms of function of NAG7 gene, a powerful new tool for the unambiguous characterization of gel separated proteins is accomplished by the combination of mass spectrometry and sequence database searching. NAG7 , a novel putative tumor suppressor gene, located on 3p25.3, was introduced into HNE1 cells by liposome transfection. We used two dimensional polyacrylamide gel electrophoresis (2 D PAGE) to identify proteins that were overexpressed in NAG7 transfected cells. After staining and image analysis, spots of interest were isolated and subjected to mass spectrometry (MS). We found nine proteins, which up regulated in NAG7 transfected cells and be identified by MS. These proteins included growth arrest specific protein, DNA binding protein, c myc promoter binding protein and caspase 6 etc., which involved in cell cycling, transcription regulation, and apoptosis. NAG7 may exert the function by up regulating the expression of these proteins.

关 键 词：NAG7基因 细胞转染 二维凝胶电泳 基质辅助激光解吸 离子化质谱 鼻咽癌相关基因 

分 类 号：Q78[生物学—分子生物学]