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SNT1与RET之间相互作用的酵母双杂交研究

Study on interaction between suc1-binding neurotrophic target 1 and RET by yeast two-hybrid method

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【作者】 矫力张勇刘翾刘秀杰张照环王永刚朱伟何成

【Author】 JIAO Li,ZHANG Yong,LIU Xuan,LIU Xiu-jie,ZHANG Zhao-huan,WANG Yong-gang,ZHU Wei,HE Cheng~*(Department of Neurobiology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433,China)

【机构】 第二军医大学基础医学部神经生物学教研室第二军医大学基础医学部神经生物学教研室 上海200433上海200433

【摘要】 目的:研究suc1-binding neurotrophic target(SNT1)与RET之间的相互作用,寻找RET受体下游的结合蛋白,以深入认识RET下游信号转导机制。方法:将RET胞内域与LexA蛋白融合作为DNA结合蛋白,分别将SNT1、SNT1PTB及SNT1ΔPTB与B42AD蛋白融合作为激活域蛋白,共转化酵母菌后,通过β-半乳糖苷酶活性检测以及氨基酸营养缺陷生长实验分析它们之间的相互作用。结果:SNT1及SNT1PTB与RET之间在酵母中存在结合,而SNT1ΔPTB不与RET结合。结论:证实RET与SNT1之间具有相互作用,SNT1的PTB结构域在介导此相互作用的过程中非常重要。

【Abstract】 Objective:To study the interaction between RET (GDNF receptor) with suc1-bingding neurotrophic target(SNT1) and search for the possible downstream substrates or regulatory proteins of RET,so as to reveal the mechanism of downstream signal transduction of RET.Methods: The RETIC(intracellular domain of RET) was fused to LexA and the product was used as a DNA-binding domain protein.SNT1,SNT1PTB or SNT1ΔPTB(SNT1 without PTB) was separately fused to B42AD and their products were used as an activation domain protein.The cotransformants were tested by β-galactosidase activity analysis and leucine medium growth analysis was used to study the interaction between SNT1 and RET.Results: Interaction between RET and SNT1 or SNT1PTB was found in yeast,but that between SNT1ΔPTB and RET was not observed.Conclusion: It is confirmed that RET can interact with SNT1 in yeast,and the interaction may be mediated by PTB domain of SNT1.

【关键词】 RETSNT1PTB酵母双杂交蛋白相互作用
【Key words】 RETSNT1PTByeast two-hybridinteraction
【基金】 国家自然科学基金(30400123,30325022,30570939);上海市青年科技启明星计划基金(05QMX1469);上海市科技发展基金(04XD14004,04DZ14005)~~
  • 【文献出处】 第二军医大学学报 ,Academic Journal of Second Military Medical University , 编辑部邮箱 ,2006年06期
  • 【分类号】Q26-33
  • 【被引频次】1
  • 【下载频次】95
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