核基质MINT N端片段(365～1084aa)相互作用蛋白的筛选及鉴定  

Screening and identification of nuclear matrix protein MINT N-terminal fragment (365-1084aa) interacting proteins

作　　者：秦红[1] 孙强[1] 周鹏[1] 李军锋[1] 韩骅[1] 罗晓星[2] 

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 [2]第四军医大学基础部药理学教研室,陕西西安710033

出　　处：《第四军医大学学报》2004年第1期1-3,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金 (30 2 0 0 1 4 8) ;国家高技术发展规划项目(0 0 1CB50 990 6)

摘　　要：目的 :确定核蛋白MINT(Msx2 interactingnucleartargetProtein)的功能及作用机制 .方法 :利用酵母双杂交的方法 ,以MINTN端第 36 5～ 1 0 84位氨基酸 (MINT F2 )为诱饵用酵母双杂交法筛选小鼠胚胎 9日的cDNA文库 .用诱饵质粒和文库先后转化酵母宿主后 ,对 1 .0× 1 0 8个酵母克隆进行营养筛选和 β 半乳糖苷酶筛选 ,获得 1 2 8个阳性克隆 .并提取质粒进行酶切鉴定 ,将获得的 4个非重复克隆并进行序列分析 .结果 :筛选出 4个与MINT F2相互作用的蛋白 ,它们分别是 :6 7ku层黏连蛋白受体 ,,2个 1 6S核糖体RNA ,精氨酰 tRNA合成酶 .结论 :成功筛选出与MINT F2相互作用的蛋白 。AIM: To identify the function and mechanism of nuclear matrix protein MINT (Msx2 interacting nuclear target protein). METHODS: Yeast two hybrid assay was used to screen proteins that interact with a fragment of MINT F2 (365-1084aa). From 1.0×10 8 yeast clones transformed with the bait plasmid and a cDNA library of 9 dpc mouse embryo,128 were positive for nutritional screening and β galactosidase assay. Restriction digestion showed 4 independent positive clones and they were analyzed by DNA sequencing, and revealed that they represent correctly fused cDNA fragments. RESULTS: Screened 4 proteins that interacting with MINT F2, which are 67 ku laminin receptor, two of 16 S ribosomal RNA, Arginyl tRNA synthetase. CONCLUSION: Succeed screening of interacting with MINT F2 proteins, and made the basement for further study of the function and mechanism of MINT.

关 键 词：核蛋白MINT MSX2 RBP—Jk NOTCH信号途径 酵母双杂交 

分 类 号：Q785[生物学—分子生物学] Q789