酵母双杂交技术筛选与hCLP46蛋白相互作用的蛋白

目的：利用酵母双杂交技术，筛选人类白细胞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，深入探讨hCLP46的功能，为骨髓增生异常综合症MDS-AML的病理提供线索。方法：以pGBKT7-hCLP46为诱饵质粒筛选人类白细胞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：从人类白细胞cDNA文库中得到86个阳性克隆，经过验证，最终得到5个阳性克隆。对验证后的阳性克隆的外源片断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。

Identification of proteins interacting with hCLP46 from Human leukocyte cDNA Library using yeast two-hybrid system

Objective To screen the interactive proteins of hCLP46 and provide clues for thestudies on hCLP46 function and pathogenesis of MDS-AML.Methods The human leukocyte cDNA library was screened with with pGBKT7-hCLP46 as bait plasmid by yeast two hybrid system , and Leu+, Mel+, Trp+, His+ positive yeast clones were obtained , The inserted fragments of identified positive clones were sequenced and analyzed with Blast in NCBI. Results There are 86 positive yeast clones which are obtained through screening the human leukocyte cDNA library with pGBKT7-hCLP46 as bait plasmid. Four different candidate gene sequences are obtained by Blast analysis in NCBI. Conclusion Four gene sequences were obtained , these coding proteins can interact with hCLP46 ,which may be involved with pathogenesis of MDS-AML.