摘要
收集水牛孤雌激活后2细胞期早期胚胎500枚，利用0．1％链酶蛋白酶去除透明带后提取蛋白质，并使用胰蛋白酶进行酶解处理，再将酶解得到的多肽混合物经强阳离子交换色谱进行顸分离，预分离后所收集的馏分通过液相色谱串联质谱（LC—MS／MS）技术进行质谱鉴定，原始数据通过数据库搜索获得该时期蛋白质表达谱0通过SE-QUEST检索共得到778种蛋白，对其进行生物信息学初步分析，包括G0富集分析和蛋白互作网络分析等。结果发现，这些蛋白质主要参与氧化还原、蛋白质折叠、细胞内蛋白质运输等生物学过程，主要定位于线粒体和细胞膜上。本研究为后续以孤雌激活为研究模型探讨哺乳动物早期胚胎发育和母源性蛋白质调控机制提供了新的研究手段和方法。
The two-cell stage early embryo of buffalo is the research object. 500 two-cell stage early embryo of buffalo after parthenogenetic activation were collected, and the zona pellucida was removed by using 0. 1% pronase and then proteins were extracted then hydrolyzed by trypsin, the peptide mixture was separated by using the strong cation exchange chromatographic, the fraction after separation was indentified by using liquid chromatography-tandem mass spectrometry（LCMS/MS）technology for mass spectrometry, and the raw data was searched by the database then the protein expression profile of this period was done. A total of 778 protein by using SEQUEST search were obtained and then made preliminary bioinformatics analysis for this results. The results showed that these proteins were primarily involved in the oxidation and reduction processe, protein folding,intracellular protein transport and other biological processes, mainly in the mitochondria and the cell membrane. The research provided new research tools and methods for the study on parthenogenetic activation model of mammalian early embryonic and maternal protein regulation mechanism.
作者
陈富美
濮黎萍
付强
王焕景
张鹏飞
候振
徐壮壮
黄凤玲
陆阳清
张明
CHEN Fu-mei;PU Li-ping;FU Qiang;WANG Huan-jing;ZHANG Peng-fei;HOU Zhen;XU Zhuang-zhuang;HUANG Feng-ling;LU Yang-qing;ZHANG Ming(State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources , Animal Reproduction Institute, Guangxi University, Nanning 530004, Chin)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2018年第6期1237-1243,共7页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31460603)
广西高等学校高水平创新团队及卓越学者计划资助项目(31460603)
广西研究生教育创新计划博士研究生创新资助项目(YCBZ2017005)
关键词
孤雌激活
早期胚胎
蛋白质表达谱
生物信息学
parthenogenetic activation
early embryos
protein profile
bioinformatics
作者简介
陈富美（1990-），女，博士。;通讯作者，E-mail：mingzhang@gxu．edu．cn
