白鲢鱼肌原纤维蛋白双向电泳分析体系的建立  

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

作　　者：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 郁晓君 YI Shumin;LI Ruizhi;CHEN Yang;LI Jianrong;LI Xuepeng;XU Yongxia;LI Chun;DING Haochen;YU Xiaojun(College of Food Science and Engineering, Bohai University, Jinzhou 121013, China;College of Mathematics and Physics, Bohai University, Jinzhou 121013, China;Liaoning Anjoy Food Co. Ltd., Anshan 114100, China)

机构地区：[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　　处：《食品科学》2017年第1期41-46,共6页Food Science

基　　金：国家自然科学基金面上项目(31571868);辽宁省教育厅重点实验室基础研究项目(LZ2015002;LZ2014047);"十二五"国家科技支撑计划项目(2015BAD17B03);辽宁省高等学校创新团队项目(LT2014024)

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p H值梯度预制胶条p H 4~7,上样量120μg,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。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 immobilizedpH gradient (IPG) strip were selected as the main factors affecting 2-DE in this paper. We loaded 120 μg of protein onto the17 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 heldfor 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.

关 键 词：白鲢鱼 肌原纤维蛋白 双向电泳 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]