人胎脑中hSNF5结合蛋白的筛选、克隆及分析  

Screening? Cloning? and Analyzing for hSNF5 Binding Proteins inHuman Fetal Brain

作　　者：张怡[1] 杨珺[1] 吴宁华[1] 沈珝琲[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《中国医学科学院学报》2004年第5期529-532,共4页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(30028011;39930050)资助~~

摘　　要：目的寻找在神经细胞中与染色质重塑复合物中hSNF5亚单位相结合的靶蛋白并探索其功能,为阐明hSNF5在细胞中参与染色质重塑复合物对靶基因调控的机制提供线索。方法以hSNF5为诱饵,采用酵母双杂交系统筛选人胎脑MATCHMAKERcDNA文库。测定核苷酸序列,计算机软件分析核苷酸的同源性及功能结构域。结果获得9个阳性克隆。DNA序列分析及同源性比较表明,1个克隆的序列与推导蛋白FLJ20643羧基端的mRNA同源性高达97%,4个克隆与GenBank中可编码蛋白的序列有较高的同源性,其他4个克隆编码的氨基酸顺序与GenBank中的已知序列无同源性。结论在人胎脑cDNA文库中克隆到hSNF5的结合蛋白,这些蛋白质可能通过hSNF5募集染色质重塑复合物以调节其靶基因的转录活性熏及其参与的细胞功能。Objective To identify novel binding proteins of hSNF5? a subunit of chromatin remodeling complex in human fetal brain. Methods The yeast two-hybrid system was used for this study. Positive cDNA clones were sequenced. Sequence homology and putative functional domains were analyzed and compared with databank. Results Nine positive clones obtained were analyzed? among which the sequence of one clone was 97% homologous to the 3' mRNA of a hypothetical protein FLJ20643? while other four clones were related to protein coding sequences existed in the GenBank. The rest four clones were not in frame with any known protein coding sequence. Conclusions Clones encoding for hSNF5 binding protein exists in cDNA library of human brain. These proteins may recruit chromatin remodeling complex via hSNF5 to modulate the transcription of their target gene and the related cell functions.

关 键 词：hSNF5 酵母双杂交体系 基因调控 

分 类 号：Q51[生物学—生物化学]