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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

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)

CLC Number: