酵母双杂交筛选与果蝇C(2)M相互作用的蛋白

doi:10.16288/j.yczz.15-118

遗传 ›› 2015, Vol. 37 ›› Issue (11): 1160-1166.doi: 10.16288/j.yczz.15-118

酵母双杂交筛选与果蝇C(2)M相互作用的蛋白

岳珊珊^1,2,夏来新¹

1. 安徽大学生命科学学院,安徽 230601;
2. 中国科学院动物研究所,计划生育生殖生物学国家重点实验室,北京 100101

收稿日期:2015-03-22 修回日期:2015-08-25 出版日期:2015-11-20 发布日期:2015-09-11
通讯作者: 夏来新,教授,博士生导师,研究方向：RNA代谢和干细胞。E-mail: xialx@ioz.ac.cn E-mail:yueshanshanyss@163.com
作者简介:岳珊珊,硕士研究生,专业方向：RNA代谢和干细胞。E-mail: yueshanshanyss@163.com
基金资助:
SKLRB青年杰俊科研创新主任基金项目资助

Identification of C(2)M interacting proteins by yeast two-hybrid screening

Shanshan Yue^1,2,Laixin Xia¹

1. School of Life Sciences, Anhui University, Anhui 230601, China;
2. State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China

Received:2015-03-22 Revised:2015-08-25 Online:2015-11-20 Published:2015-09-11

摘要/Abstract

摘要： 同源染色体联会时形成的联会复合体(Synaptonemal complex, SC)是由减数分裂前期Ⅰ多种蛋白质聚集而成的超级复合结构。生殖细胞特异性的核蛋白C(2)M(Crossover suppressor on 2 of Manheim)在染色体上高度聚集可以诱导SC的形成。本文采用酵母双杂交方法,利用C(2)M的诱饵表达载体筛选果蝇cDNA文库,共发现40个可能与C(2)M相互作用的蛋白,包括多种DNA及组蛋白结合蛋白、ATPase、转录调节因子。从筛选的结果中,选取wech和Psf1基因构建了转基因果蝇,并在生殖细胞中进行了基因沉默,结果显示联会复合体的消失受到延迟。上述结果表明Wech和Psf1蛋白可能与C(2)M形成复合物,共同参与联会复合体的形成或其稳定性的维持。

关键词: 减数分裂, SC, C(2)M, 酵母双杂交, 蛋白相互作用

Abstract: The synaptonemal complex (SC) is a huge structure which assembles between the homologous chromosomes during meiotic prophase I. Drosophila germ cell-specific nucleoprotein C(2)M clustering at chromosomes can induce SC formation. To further study the molecular function and mechanism of C(2)M in meiosis, we constructed a bait vector for C(2)M and used the yeast two-hybrid system to identify C(2)M interacting proteins. Forty interacting proteins were obtained, including many DNA and histone binding proteins, ATP synthases and transcription factors. Gene silencing assays in Drosophila showed that two genes, wech and Psf1, may delay the disappearance of SC. These results indicate that Wech and Psf1 may form a complex with C(2)M to participate in the formation or stabilization of the SC complex.

Key words: meiosis, SC, C(2)M, yeast two-hybrid system, protein interaction

岳珊珊,夏来新. 酵母双杂交筛选与果蝇C(2)M相互作用的蛋白[J]. 遗传, 2015, 37(11): 1160-1166.

Shanshan Yue,Laixin Xia. Identification of C(2)M interacting proteins by yeast two-hybrid screening[J]. HEREDITAS(Beijing), 2015, 37(11): 1160-1166.

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酵母双杂交筛选与果蝇C(2)M相互作用的蛋白

Identification of C(2)M interacting proteins by yeast two-hybrid screening

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