牦牛卵泡液差异蛋白质组双向电泳图谱的构建与质谱分析  

Establishment of Two-dimensional Electrophoresis Mapping and Mass Spectrometry Analysis of the Differential Proteome of Yak Follicular Fluid

作　　者：郭宪[1] 裴杰[1] 褚敏[1] 王宏博[1] 丁学智[1] 阎萍[1] 

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,甘肃省牦牛繁育工程重点实验室,兰州730050

出　　处：《中国畜牧兽医》2015年第11期3037-3043,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31301976);现代农业(肉牛牦牛)产业技术体系建设专项(CARS-38);国家科技支撑计划项目子课题(2013BAD16B09);中国农业科学院创新工程项目(CAAS-ASTIP-2014-LIHPS-01)

摘　　要：从蛋白质水平了解牦牛季节性繁殖规律,利用双向电泳与质谱鉴定技术分析牦牛卵泡液与血浆蛋白质组分变化。以青海高原牦牛卵泡液与血浆为研究对象,采用双向电泳技术构建牦牛卵泡液与血浆蛋白质双向电泳图谱,银染后利用Image Master 2DPlatinum软件分析并采用MALDI-TOF-MS进行质谱鉴定。用试剂盒ProteoExtract Albumin/IgG Removal Kit去除高丰度蛋白质后,利用2-DE技术获得了分辨率较高的卵泡液与血浆蛋白质电泳图谱,卵泡液与血浆蛋白质图谱对比分析共发现了24个差异表达蛋白质点,其中2个蛋白质点表达上调,22个蛋白质点表达下调。经质谱分析,最终成功鉴定出8个蛋白质点、5个未知蛋白质点。本研究成功构建了蛋白质图谱及分离鉴定的差异蛋白质,为从蛋白质水平揭示牦牛卵泡发育规律及了解卵母细胞发育的微环境提供了试验依据。In order to know the law of seasonal reproduction of yak（Bos grunniens）at protein level,the composition changes of follicular fluid and plasma protein were analyzed by two-dimensional electrophoresis（2-DE）and mass spectrometry（MS）.The study employed Qinghai plateau yak follicular fluid and plasma as the research objects.The maps were obtained by 2-DE.After silver staining,the differential proteome was analyzed using the Image Master 2DPlatinum software and MALDI-TOF-MS（matrix-assisted laser desorption ionization time of flight mass spectrometry）.First we described the purification of follicular fluid and plasma samples using ProteoExtract Albumin/IgG Removal Kit,and then described the maps of protein from these purified follicular fluid and plasma using the 2-DE system.24 different expression protein spots were found with Image master 2Dplatinum soteware,2of these proteins were up-regulated,and 22 of these proteins were down-regulated in yak follicular fluid.Through MALDI-TOF-MS identification,8protein spots were successfully identified and 5protein spots were unknown in NCBI_Bos tarus.Establishment of 2-DE maps,isolation and identification of differential protein were successfully to pro-vide experimental basis for revealing the law of follicular development and developmental microenviroment of oocytes at protein level in yak.

关 键 词：牦牛 卵泡液 血浆 差异蛋白质组 双向电泳 质谱鉴定 

分 类 号：Q591.2[生物学—生物化学]