食品科学

• 基础研究 • 上一篇    下一篇

白鲢鱼肌原纤维蛋白双向电泳分析体系的建立

仪淑敏1,李睿智1,陈 杨1,励建荣1,*,李学鹏1,徐永霞1,李 春2,丁浩宸3,郁晓君3   

  1. 1.渤海大学食品科学与工程学院,辽宁 锦州 121013;2.渤海大学数理学院,辽宁 锦州 121013;3.辽宁安井食品有限公司,辽宁 鞍山 114100
  • 出版日期:2017-01-15 发布日期:2017-01-16

Establishment of Two-Dimensional Gel Electrophoresis System for Myofibrillar Proteins of Silver Carp (Hypophthalmichthys molitrix)

YI Shumin1, LI Ruizhi1, CHEN Yang1, LI Jianrong1,*, LI Xuepeng1, XU Yongxia1, LI Chun2, DING Haochen3, YU Xiaojun3   

  1. 1. College of Food Science and Engineering, Bohai University, Jinzhou 121013, China;
    2. College of Mathematics and Physics, Bohai University, Jinzhou 121013, China;
    3. Liaoning Anjoy Food Co. Ltd., Anshan 114100, China
  • Online:2017-01-15 Published:2017-01-16

摘要: 以白鲢鱼为研究对象,建立并优化其肌原纤维蛋白的双向电泳技术。结果表明,选用固相pH值梯度预制胶条pH 4~7,上样量120 μg,等电聚焦程序Ⅲ(50 V主动水化15 h,250 V、2 h,1 000 V、2.5 h及4 000 V、3 h三段式除盐,8 000 V、2 h和10 000 V、1.5 h两段式升压,10 000 V聚焦80 000 V·h,500 V保持10 h)以及12%的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离胶条件下进行双向电泳,凝胶银染后用PDQuest软件分析,得到具有高分离度的肌原纤维蛋白双向电泳图谱,蛋白点清晰。

关键词: 白鲢鱼, 肌原纤维蛋白, 双向电泳

Abstract: A two-dimensional gel electrophoresis (2-DE) system for the separation of myofibrillar proteins from silver carp
was established and optimized in this study. The key factors of isoelectric focusing (IEF) and the pH gradient of immobilized
pH gradient (IPG) strip were selected as the main factors affecting 2-DE in this paper. We loaded 120 μg of protein onto the
17 cm pH 4-7 IPG strip, and the IEF program III was as follows: 50 V for 15 h, three-stage desalting involving 250 V for 2 h,
1 000 V for 2.5 h and 4 000 V for 3 h, 8 000 V for 2 h and 10 000 V for 1.5 h, and 10 000 V for 80 000 V·h and 500 V held
for 10 h. The gel concentration was 12%. The gel was stained with sliver nitrate, scanned and analyzed using PDQuest
software. A high-resolution 2-DE map for myofibrillar proteins from silver carp was obtained finally.

Key words: silver carp, myofibrillar protein, two-dimensional gel electrophoresis (2-DE)

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