中肋骨条藻蛋白质双向电泳样品制备方法的比较

海洋学研究 ›› 2012, Vol. 30 ›› Issue (4): 72-77.

中肋骨条藻蛋白质双向电泳样品制备方法的比较

王秀秀, 陈纪新, 黄邦钦^*

福建省陆海界面生态环境重点实验室,厦门大学环境与生态学院,福建厦门 361005

收稿日期:2012-08-12 修回日期:2012-08-26 出版日期:2012-12-15 发布日期:2022-11-30
通讯作者: *黄邦钦,教授, E mail: bqhuang@xmu.edu.cn
作者简介:王秀秀(1982-),女,河北张家口市人,博士研究生,主要从事浮游植物分子生态学研究。E-mail: xiuxiuwang0420@126.com
基金资助:
国家自然科学基金资助项目(40576055); 国家海洋公益性行业科研专项经费资助项目(201105021)

Comparison of protein sample preparation methods of two-dimensional electrophoresis for Skeletonema costatum

WANG Xiu-xiu, CHEN Ji-xin, HUANG Bang-qin^*

Fujian Provincial Key Laboratory of Coastal Ecology and Environmental St udies, Collegeof the Environment and Ecology, Xiamen University, Xiamen 361005, China

Received:2012-08-12 Revised:2012-08-26 Online:2012-12-15 Published:2022-11-30

摘要/Abstract

摘要： 比较了裂解液法和直接三氯乙酸(TCA)-丙酮沉淀法对中肋骨条藻Skeletonema costatum 蛋白双向电泳的提取效果并优化了提取条件,结果表明:裂解液-TCA-丙酮沉淀法在去除胞内干扰物质方面,较裂解液-超滤管法和裂解液-试剂盒法的效果都好。裂解液-TCA-丙酮沉淀法和直接TCA-丙酮沉淀法都能取得背景干净、蛋白点清晰的双向电泳图谱,但后者的双向电泳图谱聚焦更完全,在进一步优化条件后(即蛋白沉淀12 h并增加超声波洗涤过程),可作为提取中肋骨条藻蛋白的最适方法。

关键词: 中肋骨条藻, 蛋白质组学, 双向电泳, 蛋白提取方法

Abstract: Lysis buffer method and trichloroacetic acid ( TCA)-acetone precipitation method both are the common methods in algal protein abstraction. We compared the 2-DE maps of these two protein extraction methods to find out the best protocol for Skeletonema costatum. As a result,the Lysis buffer-TCA-acetone method had better effect than the other two lysis buffer methods (lysis buffer-microcon &. lysis buffer-2-D clean-up kit) in removing the intracellular interferential factors, such as salt, nucleic acid, polyphenol and polysaccharide. The lysis buffer-TCA-acetone method and straight TCA-acetone precipitation method both represented clean background and clear protein dots in 2-DE maps while the later method showed better isoelect rofocusing results. We optimized the straight TCA-acetone precipitation methods. The TCA-acetone precipitation with 12 hours precipitation and a following ultrasonic cleaning process method was confirmed to be the appropiate sample preparation method for S. costatum for two-dimensional electrophoresis.

Key words: Skeletonema costatum, proteomics, two-dimensional electrophoresis, protein extraction method

中图分类号:

Q946

王秀秀, 陈纪新, 黄邦钦. 中肋骨条藻蛋白质双向电泳样品制备方法的比较[J]. 海洋学研究, 2012, 30(4): 72-77.

WANG Xiu-xiu, CHEN Ji-xin, HUANG Bang-qin. Comparison of protein sample preparation methods of two-dimensional electrophoresis for Skeletonema costatum[J]. Journal of Marine Sciences, 2012, 30(4): 72-77.

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