酵母双杂交技术筛选并回转验证在胞内与PML-C结构域相互作用的蛋白  

Screening and Identification of Target Proteins Interacting with Structural Domain of PML-C by Yeast Two-Hybrid System Techniques

作　　者：朱丹[1] 王翀 刘北忠[1] 钟梁[1] 王春光[1] 吴燕[1] 

机构地区：[1]重庆医科大学临床检验诊断学教育部重点实验室,重庆市400016

出　　处：《医学分子生物学杂志》2010年第3期242-246,共5页Journal of Medical Molecular Biology

基　　金：国家自然科学基金（No.30300449）,国家中医药管理局科研基金（No.02-03ZP52）

摘　　要：目的 利用酵母双杂交技术在活细胞内筛选并回转验证与PML-C结构域相互作用的蛋白质.方法 通过诱饵质粒pGBKT7-PML-C,利用酵母双杂交系统从白血病细胞cDNA文库中筛选与PML-C结构域相互作用的蛋白质.结果 利用酵母双杂交技术筛选到43个能与PML-C结构域相互作用的克隆;经进一步的归类与酵母回转试验得到9个阳性克隆.结论 在细胞内PML-C结构域能与多种蛋白质有相互作用.中性粒细胞弹性蛋白酶（neutrophil elastase,NE）介导的急性早幼粒细胞白血病的发生可能与这些相互作用所致的生物学功能改变有关.Objective Screen and identify protein interacting with the domain of PML with coiled-coil structure （PML-C） via yeast two-hybrid techniques. Methods The bait vector pGBKT7-PML-C was constructed for screening proteins interacting with PML-C in leukemia cell cDNA expression library via yeast two-hybrid system. Results Forty-three proteins were screened to interact with PML-C by yeast two-hybrid system, nine positive clones were identified by retransformation in yeast. Conclusions There were some intracellular proteins interacting with PML-C. The pathogenesis mediated by NE of leukemia may be related to biological function altered by certain proteinprotein interactions.

关 键 词：PML—C 酵母双杂交技术 蛋白质相互作用 

分 类 号：R733[医药卫生—肿瘤]