拟穴青蟹卵巢蛋白质双向电泳的样品制备方法  

Preparation Methods of Ovary Protein Samples in Green Mud Crab Scylla paramamosain for Two-dimensional Electrophoresis

作　　者：王为刚[1] 王艺磊[1] 张子平[2] 冯建军[1] 陈锦民[1] 邹志华[1] 谢芳靖[1] 林鹏[1] 

机构地区：[1]集美大学水产学院水产科学技术与食品安全省高校重点实验室,福建厦门361021 [2]德克萨斯州立大学化学和生物化学系,美国圣马克斯tx78666

出　　处：《水产科学》2011年第6期352-355,共4页Fisheries Science

基　　金：国家自然科学基金资助项目(31072200);福建省高等学校新世纪优秀人才支持计划项目(闽教科[2007]20号);福建省科技厅高校专项基金资助项目(2008F5039);集美大学创新团队基金资助项目(2010A001)

摘　　要：为探索适用于拟穴青蟹卵巢蛋白质双向电泳的样品制备方法,比较了TCA/丙酮沉淀法和3种裂解液抽提法对总蛋白的提取效果,4种方法分别得到133、369、306和417个蛋白点。采用裂解液Ⅲ(7 mol/L尿素、2 mol/L硫脲、4% CHAPS、40 mmol/L Tris-base、65 mmol/L DTT、2% IPG Buffer、Protease in hibitor cocktail、1 mmol/L EDTA)提取蛋白所得图谱效果最好,背景清晰,蛋白点最多。The ovary proteomics were comparatively analyzed in green mud crab Scylla paramamosain by four methods , i. e. trichloroacetic acid/acetone precipitation and three different lysis buffers extraction in order to develop an efficient protein preparation method suitable for ovary proteomics anal, ysis. The results showed that 133, 369, 306 and 417 protein spots were detected on 2-DE gels in the four methods. The lysis buffer Ⅲ （7 mol/L urea, 2 mol/L thiourea, 4% CHAPS, 40 mmol/L Tris-base, 65 mmol/L DTT, 2% IPG Buffer, Protease inhibitor cocktail, and 1 mmol/L EDTA） extraction was found to be the most appropriate for proteomics analysis on account of the highest resolution, and the most protein spots.

关 键 词：拟穴青蟹 卵巢 蛋白质组 双向电泳 样品制备 

分 类 号：Q81[生物学—生物工程]