水产学报2011，Vol.35Issue(9)：1362-1368,7.DOI:10.3724/SP.J.1231.2011.17509
坛紫菜叶状体蛋白质双向电泳样品制备方法的比较
Comparison of protein preparation methods of Porphyra haitanensis thalli for two-dimensional electrophoresis
摘要
Abstract
Protein preparation is a key step in two-dimensional electrophoresis(2-DE) analysis. In order to establish a suitable protein preparation method for Porphyra haitanensis thalli, three sample preparation methods of plant, trichloroacetic acid/acetone precipitation ( TCA/ACE) method, urea/thiourea extraction (UREA/THI) method and phenol extraction methanol/ammonium acetate precipitation method, and three protein lysis buffers were compared. The results showed that the optimal method of proteins preparation of P. Haitanensis thalli was extracted using TCA/ACE method and dissolved with the lysis buffer C(7 mol/L urea,2 mol/L thiourea,2%CHAPS,2%TritonX-100,2% IPG Buffer pH 3-10,65 mmol/L DTT). Using this method,the yield was the highest and 2-DE map has the most spots,clear backgrounds,high resolution and great repeatability. At the same time, we also found that using the IPG strips (pH 4 -7) in 2-DE analysis can extract the protein of P. Haitanensis thalli better than IPG strips (pH 3-10).关键词
坛紫菜/蛋白质组/双向电泳/样品制备Key words
Porphyra haitanensis/proteome/two-dimensional electrophoresis/sample preparation分类
农业科技引用本文复制引用
杭楠,谢潮添,陈昌生,纪德华,徐燕..坛紫菜叶状体蛋白质双向电泳样品制备方法的比较[J].水产学报,2011,35(9):1362-1368,7.基金项目
国家自然科学基金项目(40806065) （40806065）
海洋公益性行业科研专项(201105008) （201105008）
公益性行业(农业)科研专项(200903030) （农业）
福建省杰出青年基金项目(20L0J06016) （20L0J06016）
福建省教育厅新世纪优秀人才项目(JA10186) （JA10186）
