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金鱼胚胎蛋白质双向凝胶电泳方法建立及优化
引用本文:喻小燕,陈妍,曹晶,谢锦云. 金鱼胚胎蛋白质双向凝胶电泳方法建立及优化[J]. 金陵科技学院学报, 2009, 25(2): 91-96
作者姓名:喻小燕  陈妍  曹晶  谢锦云
作者单位:1. 湖南信息职业技术学院,湖南,长沙,410200;湖南师范大学生命科学学院,湖南,长沙,410081
2. 湖南师范大学生命科学学院,湖南,长沙,410081
摘    要:以金鱼胚胎为材料,以不同样品制备方式、裂解液成分、固相pH梯度胶条的选择、凝胶浓度为侧重点,建立并优化金鱼胚胎总蛋白质组最佳双向凝胶电泳技术条件.研究表明,采用匀浆器匀浆含硫脲和NP-40裂解液提取蛋白,应用pH值为3~10的非线性IPG胶条和浓度14%的SDS-PAGE凝胶进行双向电泳,经优化电泳电流、时间等参数后,获得了重复性和分辫率理想的金鱼胚胎总蛋白质双向电泳图谱,为金鱼胚胎蛋白质组分析莫定了基础,为揭示金鱼单倍体发育异常的机制提供了技术保障.

关 键 词:金鱼胚胎  蛋白质组  双向凝胶电泳

Establishment and Optimization of Two-dimensional Gel Electrophoresis Condition for Goldfish Embryos Proteome
YU Xiao-yan,CHEN Yan,CAO Jing,XIE Jin-yun. Establishment and Optimization of Two-dimensional Gel Electrophoresis Condition for Goldfish Embryos Proteome[J]. Journal of Jinling Institute of Technology, 2009, 25(2): 91-96
Authors:YU Xiao-yan  CHEN Yan  CAO Jing  XIE Jin-yun
Affiliation:1.Hunan College of Information;Changsha 410200;China;2.Hunan Normal University;Changsha 410081;China
Abstract:To develop and optimize a two-dimensional(2-DE) gel electrophoresis condition for goldfish embryos proteome,total protein of goldfish embryos is extracted,and the optimal conditions for sample preparation and 2-DE are screened by comparing the following parameters: embryos broken methods,contents of lysis buffer,the concentration of gel,and linear or non-linear immobilized pH gradient gels.The results demonstrate that a satisfactory 2-DE map with good repetition and resolution is obtained and suitable condi...
Keywords:goldfish  embryos proteome  two-dimensional gel electrophoresis  
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