绵羊BMPR1B基因真核表达及产物互作蛋白的鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2019-0447

摘要/Abstract

摘要： 旨为研究BMPR1B蛋白及其相互作用蛋白在绵羊卵巢卵母细胞发育及排卵的作用通路。通过构建BMPR1B基因真核表达体系并表征BMPR1B蛋白质,利用CoIP-MS技术鉴定母羊卵巢提取物中与BMPR1B特异性相互作用的蛋白质,构建目的蛋白质互作网络,通过生物信息学分析并预测BMPR1B蛋白及其相互作用蛋白在绵羊卵巢卵母细胞发育及排卵的作用通路。母羊卵巢提取物中23种蛋白质与BMPR1B蛋白具有关联性,其中6种目的蛋白（BMP2、BMP4、GDF5、GDF9、RhoD和HSP10）与BMPR1B具有特异性相互作用,通过生物信息学分析表明目的蛋白在TGF-β信号转导通路、卵巢类固醇生成通路和MAPK信号转导通路构成复杂且紧密的交互网络,基于互作结果设计BMPR1B蛋白调节绵羊卵母细胞的发育和排卵的代谢途径。BMPR1B蛋白与绵羊产羔性状途径中Smads家族蛋白具有交互作用,在绵羊卵母细胞发育和排卵中具有重要作用。

关键词: 绵羊, BMPR1B, 互作蛋白, CoIP-MS, 生物信息学

Abstract: This work aims to study the roles and pathways of BMPR1B protein and its interactive protein in the development and ovulation of sheep ovarian oocyte. By constructing the eukaryotic expression system of BMPR1B gene and characterizing BMPR1B protein,CoIP-MS technology was used to identify the specifically interactive proteins with BMPR1B in ewe ovarian extract,and the target protein interaction network was established. Then we used bioinformatics to analyze and predict the interaction pathways of BMPR1B protein and its interaction proteins in the development and ovulation of sheep ovarian oocyte. There were 23 proteins from ewes ovary extract that were correlated with BMPR1B,while 6 target proteins（BMP2,BMP4,GDF5,GDF9,RhoD and HSP10）specifically interacted with BMPR1B. Bioinformatics analysis showed that the target protein constituted complex and close interactions with TGF-β signal transduction pathway,ovarian steroid generation pathway and MAPK signal transduction pathway,and the metabolic pathways of BMPR1B protein regulating the development and ovulation of sheep oocytes was designed based on the results of their interactions. BMPR1B protein interacts with Smads family proteins in lambing trait pathway and plays an important role in the development and ovulation of sheep oocyte.

Key words: ewes, BMPR1B, protein-protein interactions, CoIP-MS, bioinformatics

贾建磊, 陈倩, 靳继鹏, 袁赞, 张利平. 绵羊BMPR1B基因真核表达及产物互作蛋白的鉴定[J]. 生物技术通报, 2019, 35(12): 94-104.

JIA Jian-lei, CHEN Qian, JIN Ji-peng, YUAN Zan, ZHANG Li-ping. Eukaryotic Expression of BMPR1B in Sheep and Identification of Its Interaction Proteins[J]. Biotechnology Bulletin, 2019, 35(12): 94-104.

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