从小鼠11.5d胚胎cDNA文库中筛选Dishevelled2相互作用蛋白  

Screening of Proteins Interacting with Dishevelled2 in Mouse 11.5dpc Embryo Library

作　　者：黄世思[1] 翟永功[2] 韩亮[1] 张新军[1] 王银银[1] 常智杰[1] 

机构地区：[1]人类基因组研究所,清华大学生物科学与技术系,清华大学生命科学与医学研究院,北京100084 [2]北京师范大学,生命科学学院,生物医学研究所,北京100875

出　　处：《生物工程学报》2004年第5期750-753,共4页Chinese Journal of Biotechnology

基　　金：国家杰出青年科学基金海外青年学者合作研究基金 (No .3 0 2 2 80 0 7); 863项目 (No .2 0 0 4AA2 2 10 60 )资助~~

摘　　要：Dishevelled (Dvl)是个多功能、进化上非常保守的蛋白 ,在Wnt信号传导通路中起着重要的作用。为了研究Dishevelled介导Wnt信号传递的分子机制 ,利用GAL4酵母双杂交系统筛选了小鼠 1 1 5d胚胎cDNA文库 ,发现了1 5个可与小鼠Dvl2DEP结构域和羧基端相互作用的蛋白质。将阳性文库质粒测序并对测序结果做BLAST分析 ,发现其中一个阳性克隆是编码Gli3蛋白氨基端 (6 1 2 2aa)的cDNA片段 。Dishevelled proteins are multifunctional and highly conserved. These proteins are also required for the specification of cell fate and polarity by secreted Wnt proteins. To investigate the molecular mechanism of Dishevelled in mediating Wnt signal transduction, a mouse 11 5dpc embryo library was screened by yeast-two-hybrid system to find mouse Dishevelled2 DEP domain and C-terminal interacting proteins. 15 possitive clones were identified from 4 1×10 6 transformants. The DNA sequences of the positive AD/library plasmids were determined. The BLAST results revealed that one of the positive clones contained N-terminus cDNA fragments (amino acids 6-122) of Gli3 protein. The interaction between Dvl2 and Gli3 detected by yeast two-hybrid system suggests that Gli3 might play a role in some biological processes with Dishevelled.

关 键 词：小鼠 胚胎 CDNA文库 WNT信号通路 发育 酵母双杂交 

分 类 号：Q71[生物学—分子生物学] Q26