水牛卵泡液差异蛋白质双向电泳方法的建立及质谱分析

doi:10.11843/j.issn.0366-6964.2013.08.010

Abstract

Abstract:

This study was undertaken to evaluate protein expression differences between mature and immature follicular fluids of swamp buffalo in Guangxi. The research used two-dimensional gel electrophoresis (2DE) coupled to Matrix Assisted Laser Desorption/Ionization Time-of-flight Tandem Mass Spectrometry (MALDI-TOF/TOF) to analyze the different proteins, and the establishment of a suitable 2DE system for separating buffalo follicular fluid proteins was described. The results indicated that acetone precipitation in a loading volume of 350 μg in 24 cm IPG strips (pH 4-7) improved resolution and the quality of the 2DE maps, and 11 different expression protein spots were found with Image Master 2D platinum software. Compared with the mature follicular fluid 2DE map, 5 of these proteins were up-regulated in immature buffalo follicular fluid, 3 were down-regulated, 1 deleted and 2 proteins specifically expressed. Finally, 4 proteins were successfully identified by mass spectrometry; these are peroxiredoxin-2, aldose reductase, fibrinogen gamma-B chain precursor, and transthyretin precursor. In conclusion, an appropriate 2DE system for separating buffalo follicular fluid proteins was established and optimized. Different expression proteins from mature and immature buffalo follicular fluids were successfully analyzed and identified to provide new research insight into the developmental microenviroment and maturation mechanism for buffalo oocytes.

CLC Number:

S823.8⁺3 .2

HUANG Yu-lin, HUANG De-lun, GUAN Jun-liang, PAN Qiu-zhuang, FU Qiang,HUANG Feng-ling, ZHANG Ming, LU Ke-huan. Establishment of Two Dimensional Electrophoresis Method and Mass Spectrumetry Analysis of the Differential Proteins of Buffalo Follicular Fluid[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(8): 1244-1250.

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Establishment of Two Dimensional Electrophoresis Method and Mass Spectrumetry Analysis of the Differential Proteins of Buffalo Follicular Fluid

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