›› 2012, Vol. 39 ›› Issue (4): 64-67.

• 生物技术 • 上一篇    下一篇

水牛卵泡液液相色谱分离及差异蛋白质分析

付强, 杨丽, 刘振方, 张明, 卢克焕   

  1. 广西大学动物繁殖研究所,亚热带农业生物资源保护与利用国家重点实验室,广西南宁 530004
  • 收稿日期:2011-09-22 修回日期:1900-01-01 出版日期:2012-04-20 发布日期:2012-04-20
  • 通讯作者: 张明

Liquid Chromatography Separation and Differential Proteins Analysis of Buffalo Follicular Fluid

FU Qiang, YANG Li, LIU Zhen-fang, ZHANG Ming, LU Ke-huan   

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources; Animal Reproduction Institute, Guangxi University, Nanning 530004, China
  • Received:2011-09-22 Revised:1900-01-01 Online:2012-04-20 Published:2012-04-20

摘要: 本研究采用液相色谱-质谱分离鉴定技术,比较不同直径卵泡液(直径<2 mm、直径2~4 mm、直径>4 mm)的蛋白质组分差异,收集显著差异的馏分,还原烷基化处理后酶解成肽段,再经反相色谱二次分离后质谱鉴定。结果表明,在优势卵泡中(直径>4 mm)存在显著高表达的蛋白质峰,质谱鉴定结果为转铁蛋白(transferrin,TRF)。转铁蛋白的功能和表达的变化提示,该蛋白质的高表达可作为潜在排卵的标志物之一。同时,本研究采用的LC-MS技术将为卵泡液大规模蛋白质组分离和表达谱构建提供参考。

关键词: 水牛; 卵泡液; 蛋白质; 液相色谱分离; 转铁蛋白

Abstract: The proteins change in follicular fluid play an important role during oocyte maturation and ovulation. In this study, proteins from differential diameter follicular fluid (Φ<2 mm, Φ 2 to 4 mm,and Φ>4 mm) were compared using liquid chromatography and mass spectrometry (LC-MS), The differential fraction was collected and further treated including reduction, alkylation and enzyme digestion, the peptides were identified after second time liquid chromatography separation. The results indicated thatthere was a remarkable protein peak in dominant follicular fluid, the protein was transferrin (TRF) in this peak. The function and expression of TRF suggested that TRF could be an marker of potential ovulation. At the same time, LC-MS technique make the base of buffalo follicular fluid separation and proteome profile construction.

Key words: buffalo; follicular fluid; protein; liquid chromatography; transferrin

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