›› 2015, Vol. 42 ›› Issue (11): 3037-3043.doi: 10.16431/j.cnki.1671-7236.2015.11.031

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

GUO Xian, PEI Jie, CHU Min, WANG Hong-bo, DING Xue-zhi, YAN Ping   

  1. Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China
  • Received:2015-04-14 Online:2015-11-20 Published:2015-11-26

Abstract: 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 2D Platinum 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 2D platinum soteware,2 of these proteins were up-regulated,and 22 of these proteins were down-regulated in yak follicular fluid.Through MALDI-TOF-MS identification,8 protein spots were successfully identified and 5 protein spots were unknown in NCBI_Bos tarus.Establishment of 2-DE maps,isolation and identification of differential protein were successfully to provide experimental basis for revealing the law of follicular development and developmental microenviroment of oocytes at protein level in yak.

Key words: yak; follicular fluid; plasma; differential proteome; two-dimensional electrophoresis; mass spectrometry identification

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