酵母双杂交技术筛选hCLP46的相互作用蛋白  

Screening Proteins Interacting with hCLP46 from Human Leukocyte cDNA Library Using Yeast Two-Hybrid System

作　　者：张文娟[1] 杜杰[1] 常乃柏[2] 张恬[1] 王嵬[1] 刘利新[1] 

机构地区：[1]中国科学院研究生院生命科学学院,北京100049 [2]北京医院,北京100730

出　　处：《中国生物工程杂志》2008年第6期23-27,共5页China Biotechnology

基　　金：国家自然科学基金资助项目(30670889,30771193)

摘　　要：目的:利用酵母双杂交技术,筛选人白细胞cDNA文库中能与人类CD34+干/祖细胞异常表达蛋白hCLP46(human CAP10-like protein46)相互作用的未知蛋白。通过对其相互作用蛋白的筛选和研究,研究hCLP46基因在骨髓增生异常综合症MDS-AML的作用机制,为MDS-AML的临床治疗和诊断提供理论基础。方法:以pGBKT7-hCLP46为诱饵质粒筛选人白细胞cDNA文库,得到阳性克隆,并对验证后的阳性克隆的外源性片段进行测序及同源性分析。结果:经过两次筛选、验证,从人白细胞cDNA文库中得到5个阳性克隆,对验证后的阳性克隆的外源片段进行测序及同源性分析,最终得到4个不同的候选基因序列。结论:应用酵母双杂交系统,共筛选得到4个不同的基因,其编码蛋白与hCLP46有相互作用,可能与MDS-AML发病机制相关。Objective ： To screen the interactive proteins of hCLP46 for exploring the role of hCLP46 function on the pathogenesis of MDS-AML. Methods： The human leukocyte cDNA library was screened with pGBKT7- hCLP46 as bait plasmid by yeast two hybrid system. Leu＋, Mel＋, Trp＋, His＋ positive yeast clones were obtained. The exogenous fragments of identified positive clones were sequenced and analyzed with Blast in NCBI. Results：There are five positive yeast clones which were obtained through screening the human leukocyte cDNA library with pGBKT7-hCLP46 as bait plasmid. Four hCLP46-iteractive candidate genes were confirmed by DNA sequencing and Blast analysis in NCBI. Conclusion：Four gene sequences were obtained with their coding proteins interacting with hCLP46 , these candidate genes possibly involve with pathogenesis of MDS-AML.

关 键 词：酵母双杂交 骨髓增生异常综合症hCLP46 蛋白质互相作用 

分 类 号：Q819[生物学—生物工程]