食品科学 ›› 2013, Vol. 34 ›› Issue (9): 14-18.doi: 10.7506/spkx1002-6630-201309004

• 基础研究 • 上一篇    下一篇

适合蜂王浆蛋白质组的双向电泳技术

赵方圆1,2,吴亚君1,韩建勋1,陈 颖1,*,葛毅强2,3,*   

  1. 1.中国检验检疫科学研究院,北京 100123;2.中国农业大学食品科学与营养工程学院,北京 100083;
    3.中国农村技术开发中心,北京 100045
  • 收稿日期:2012-05-28 修回日期:2013-03-18 出版日期:2013-05-15 发布日期:2013-05-07
  • 通讯作者: 陈颖 E-mail:chenyingcaiq@163.com

A Two-dimensional Gel Electrophoresis Protocol for Proteomic Study of Royal Jelly

ZHAO Fang-yuan1,2,WU Ya-jun1,HAN Jian-xun1,CHEN Ying1,*,GE Yi-qiang2,3,*   

  1. 1. Chinese Academy of Inspection and Quarantine, Beijing 100123, China;
    2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China;
    3. China Rural Technology Development Center, Beijing 100045, China
  • Received:2012-05-28 Revised:2013-03-18 Online:2013-05-15 Published:2013-05-07
  • Contact: CHEN Ying E-mail:chenyingcaiq@163.com

摘要:

为建立适用于蜂王浆蛋白质组研究的双向电泳技术,对其中的IPG胶条的选择、蛋白质上样量、等电聚焦时间及SDS-PAGE凝胶浓度等关键步骤进行探索和优化。结果显示:蜂王浆蛋白质pH值主要分布在pH 5~8范围内,当上样量为150μg,聚焦时间达到32000V·h的情况下能充分地聚焦,第二向使用10%的梯度胶时获得的双向电泳图谱最佳,在优化后的蜂王浆双向电泳图谱上可检测到62±4蛋白点,重复性高,分辨率较好。优化后的双向电泳技术体系适合于蜂王浆全蛋白质的双向电泳分析。

关键词: 蜂王浆, 蛋白质组, 双向电泳, 优化

Abstract:

In order to establish a two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of royal jelly
(RJ), IPG strip with appropriate pH range, optimal loading amount for protein, focusing time of IEF, and separating gel
concentration was used. The results showed that the best 2-DE image of RJ proteins using IPG strip with pH 5—8, protein
loading amount of 150 μg, focusing time of 32000 V•h, and separating gel concentration of 10% was achieved. Moreover,
62 ± 4 protein spots were detected on the gel with high repeatability and resolution. The 2-DE system after optimization is
suitable for the analysis of RJ proteins, which provides theoretical basis for the proteomics of RJ.

Key words: royal Jelly (RJ), proteomics, two-dimensional gel electrophoresis (2-DE), optimization

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