酵母双杂交筛选NGAL相互作用蛋白  

Identification of proteins interacting with NGAL by yeast two-hybrid technique

作　　者：麦瑞琴[1] 许丽艳[1] 方王楷[2] 韩溟[1] 牛永东[2] 王子良[2] 李恩民[2] 

机构地区：[1]汕头大学医学院肿瘤病理研究室,广东汕头515031 [2]汕头大学医学院生物化学与分子生物学教研室,广东汕头515031

出　　处：《肿瘤防治杂志》2005年第4期241-247,共7页China Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金面上项目(39900069;30170428;30370641);广东省自然科学基金重点项目(37788);广东省自然科学基金面上项目(990799;010431);广东省高校自然科学研究项目(200033);广东省医学科研基金项目(A2001419);汕头大学研究与发展基金项目(L0004;L00012)

摘　　要：目的:中性粒细胞明胶酶相关脂萼蛋 白(neutrophilgelatinase assiociatedlipocalin, NGAL)功能不明,籍筛选与之相互作用蛋白, 构筑相互作用图谱,为揭示其功能提供新途径。 方法:利用蛋白质相互作用原理,应用酵母双杂 交技术,构建pGBKT7 NGAL载体,以NGAL 为诱饵,在人类骨髓cDNA文库中筛选与之相 互作用蛋白的基因。结果:通过酵母交配筛库、 一对一验证和β gal分析筛选出40个阳性克 隆;测序,利用网上数据库(http://www.ncbi. nlm.nih.gov)对序列进行比对,初步确定了31 种基因,其中11种为未知基因,20种为已知基 因。已知基因中含有编码区的有LGALS1、 MT2A、COX1、RNP24、ATP1B3、IGL、IGH VDJ、RARRES2、GYPA、NEB和NPC2。结论: 筛选得到31种可能与NGAL相互作用蛋白的 基因,这为研究NGAL功能及作用机制提供了 新途径。OBJECTIVE：The functions of NAGL are not clear at present. This study was to provide a new way to reveal the roles of NGAL protein by screening the proteins interacting with it and constructing the protein-protein interactive networks. METHODS： According to the principle of protein interactions, bait vector pGBKT7-NGAL was constructed and a pre-transformed human bone marrow cDNA library PACT2 the for the proteins that interacting with NGAL by yeast two-hybrid technique. RESULTS： After yeast mating and sequence-transformation screening and nutritional selection as well as β-gal analysis, 40 positive clones of putative interactions were obtained and 31 genes of them were identified by sequencing and utilizing of the data on line （http：//www. ncbi. nlm. nih. gov）. There were 11 putative genes and 20 known genes, including LGALS1, MT2A, COX1, RNP24, ATP1B3, IGL, IGH VDJ, RARRES2, GYPA, NEB and NPC2, CONCLUSIONS： Thirty-one proteins interacting with NGAL are obtained and an approach is offered for further exploring roles and biochemistry mechanisms of NGAL protein.

关 键 词：癌基因 基因表达 蛋白相互作用 酵母菌 DNA 互补/遗传学 

分 类 号：R735.1[医药卫生—肿瘤]