MINT蛋白(1～365氨基酸)与Vav1相互作用及其位点的验证  

Identification of interaction and exact region for interaction between N-terminal of MINT and Vav1

作　　者：董潇[1] 韩骅[1] 

机构地区：[1]第四军医大学基础部:医学遗传学与发育生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2005年第4期304-307,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金资助项目(30330550)

摘　　要：目的:明确MINT(Msx2 interactingnucleartarget protein)N 末端与Vav1之间的相互作用,进一步探讨MINT 的转录抑制调控功能及其机制.方法:以克隆有MINT F1片 段(1～365氨基酸)的pGBKT7 F1质粒作为诱饵,分别与 pACT2 V23#,pGADT7 V620,pACT2 V8900,pGADT7 SH2和 pGADT7 SH3几个克隆有Vav1不同结构域的质粒,用酵母双 杂交方法分析它们之间的相互作用.结果:pACT2 V23#与 pGBKT7 F1之间、pACT2 V8900与pGBKT7 F1之间有比较明 确的相互作用,而pACT2 V23#与pGBKT7 F2～F6、pGBKT7 F1与pGADT7 V620、pGADT7 SH2和pGADT7 SH3之间均无 相互作用.结论:①MINTN 端的F1段与Vav1发生相互作 用;②Vav1与MINT的F1段发生相互作用的区域位于其 SH2 SH3 C端结构域,而且独立的Vav1SH2或SH3 C端结构 域均不能发挥与MINT的F1段产生相互作用的功能.AIM: To study the interactions between the N-terminal of Msx2-interacting nuclear target protein (MINT) and Vav1 and to further clarify the transcriptional regulation of MINT and the mechanism of its function.METHODS: Yeast two-hybrid assay was conducted to analyze the interaction between the N-terminal of MINT and Vav1.The bait plasmid was constructed by cloning MINT-Fx (X represents 1-6) into the vector pGBKT7,several different fragments of the C-terminal of Vav1 were cloned into the vector pGADT7 or pACT2 and their interaction was tested by the growth of yeast assay.RESULTS: Definite interaction was found between pACT2-V23# and pGBKT7-F1 or pACT2-V8900 and pGBKT7-F1,while no interaction was found between other clones.CONCLUSION: ① MINT can interact with Vav1 by its N-terminal.② The exact region for the interaction of MINT and Vav1 lies in the SH2-SH3-C terminal of Vav1,but single SH2 or SH3-C terminal of Vav1 can not interact with MINT.

关 键 词：MINT Msx2 癌基因Vav 酵母菌 杂交 遗传 

分 类 号：Q344[生物学—遗传学]