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目的 确定人gp130结合分子 (GAM)与transducinlikeenhancerofsplit 1(TLE1)分子间是否存在相互作用及相互作用的结构基础 ,以深入研究这两种分子的功能。方法 扩增编码GAM与TLE1分子不同结构域的cDNA ,构建含有这些不同结构域的酵母双杂交载体 ,通过酵母双杂交系统分析和免疫共沉淀试验 ,研究这两种分子间的相互作用。结果 DNA序列测定证实成功构建了含GAM与TLE1分子不同结构域的酵母双杂交载体 ,酵母双杂交分析表明GAM与TLE1分子通过氨基端的Q结构域相互结合 ,免疫共沉淀试验证实了这一发现。结论 证实GAM与TLE1分子通过氨基端的Q结构域相互结合
Objective To determine the structural basis for the interaction and interaction between the human gp130 binding molecule (GAM) and the transducinlike enhanceroflit 1 (TLE1) molecule to further investigate the function of these two molecules. Methods cDNAs encoding different domains of GAM and TLE1 were amplified to construct yeast two-hybrid vectors containing these different domains. The interaction between the two molecules was studied by yeast two-hybrid system analysis and co-immunoprecipitation assay. Results DNA sequencing confirmed the successful construction of a yeast two-hybrid vector containing different domains of GAM and TLE1. Yeast two-hybrid analysis showed that GAM and TLE1 molecules bound to each other through the amino-terminal Q domain. The co-immunoprecipitation assay confirmed this finding. The conclusion confirms that GAM and TLE1 molecules bind to each other through the amino terminal Q domain