| 用酵母双杂交系统研究胰岛素样生长因子-1突变体与其结合蛋白-3的相互作用 |
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| 引用本文： | 丁红梅,刘宝英,杜清友,王会信,薛沿宁.用酵母双杂交系统研究胰岛素样生长因子-1突变体与其结合蛋白-3的相互作用[J].生物技术通讯,2000,11(1). |
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| 作者姓名： | 丁红梅 刘宝英 杜清友 王会信 薛沿宁 |
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| 作者单位： | 北京基础医学研究所,北京,100850 |
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| 摘 要： | 为研究胰岛素样生长因子 1(IGF1)及其突变体与IGF结合蛋白 3(IGFBP3)的相互作用 ,针对IGF1的第 3、4、15、16位氨基酸残基 ,采用定点突变的方法构建了 Y15L16 ]IGF1和 Q3A4Y15L16 ]IGF1。然后分别将IGF1/IGF1突变体和IGFBP3cDNA克隆至酵母表达载体pGBT9和pACT2中 ,利用酵母双杂交技术检测IGF1/IGF1突变体和IGFBP3之间的相互作用。结果表明用酵母双杂交系统检测IGF1与其结合蛋白的结合力是可行的 ,构建的这两个IGF1突变体与IGFBP3的结合力 ,与天然IGF1相比 ,结合力大大减小
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| 关 键 词： | 胰岛素样生长因子-1 胰岛素样生长因子结合蛋白-3 酵母双杂交 基因突变 |
Characterization of an interaction between IGF1/IGF1 mutants and IGFBP3 by using the yeast two-hybrid system |
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| Ding Hongmei,Liu Baoying,Du Qingyou,Wang Huixin,Xue Yanning.Characterization of an interaction between IGF1/IGF1 mutants and IGFBP3 by using the yeast two-hybrid system[J].Letters in Biotechnology,2000,11(1). |
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| Authors: | Ding Hongmei Liu Baoying Du Qingyou Wang Huixin Xue Yanning |
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| Abstract: | In order to study the interaction between IGF1/IGF1 mutants and IGFBP3. IGF1 mutants of IGF1 and Q3 A4 Y15 L16]IGF1 were obtained with altered sites in vitro mutagenesis system. IGF1/IGF1 mutants and IGFBP3 cDNAs were inserted into the yeast expression vectors pGBT9 and pACT2 alternatively. We used the two-hybrid assay of protein-protein interaction in the yeast Y153 to study the interaction between IGF1/IGF1 mutants and IGFBP3. The results demonstrated that the two-hybrid assay provide a facile means by which to pursue a detailed understanding of the interaction between IGF1/IGF1 mutants and IGFBP3. We also showed that IGF1 mutants had low binding affinity with IGFBP3 compared with natives. |
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| Keywords: | IGF1 IGFBP3 yeast two-hybrid gene mutation |
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