论文部分内容阅读
结肠腺瘤性息肉病基因 (adenomatouspolyposiscoli,APC)的突变导致家族性结肠息肉腺瘤病和散发性结肠癌 ,APC基因编码一个具有多个结构域、多种磷酸化状态的大分子蛋白质 .APC蛋白可通过C段直接或间接与微管结合 ,同时还可以通过中段与微管结合 ,但其结合的机制目前还不清楚 .为进一步研究APC与其他蛋白质的相互作用 ,利用酵母双杂交技术运用APC中段 (15 0 0bp～ 4 80 0bp)构建诱饵质粒 ,筛选人胎脑cDNA文库 ,得到一个与APC相互作用的蛋白SMAP/KAP3,SMAP/KAP3是驱动蛋白KIF3A/ 3B的相关蛋白 .通过免疫共沉淀和双色免疫荧光共定位的方法 ,证实了APC与SMAP/KAP3在体内的相互作用 ,提示APC可能通过SMAP/KAP3 KIF3A/B参与沿微管的运动
The mutation of adenomatous polyposis coli (APC) leads to familial colon polyadenoma and sporadic colon cancer, and the APC gene encodes a multi-domain, multi-phosphorylated macromolecular protein.APC protein Which can be directly or indirectly linked to microtubules through C segment and can also be combined with microtubules through the middle segment.However, the mechanism of its binding is unclear.To further study the interaction between APC and other proteins, using yeast two-hybrid technique, APC The bait plasmids were constructed from the middle part (15 0 0 bp ~ 4 80 0 bp), and the human fetus brain cDNA library was screened to obtain a protein SMAP / KAP3 interacting with APC. SMAP / KAP3 is a related protein of kinesin KIF3A / 3B. And two-color immunofluorescence co-localization method confirmed the in vivo interaction of APC with SMAP / KAP3, suggesting that APC may be involved in microtubule movement through SMAP / KAP3 KIF3A / B