茶树蛋白质双向电泳样品制备技术研究

doi:10.13305/j.cnki.jts.2011.03.014

茶叶科学 ›› 2011, Vol. 31 ›› Issue (3): 173-178.doi: 10.13305/j.cnki.jts.2011.03.014

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茶树蛋白质双向电泳样品制备技术研究

李勤^1,2, 黄建安^1,2, 刘硕谦¹, 李娟^1,2, 刘仲华^1,2*

1. 国家植物功能成分利用工程技术研究中心,湖南长沙 410128;
2. 茶学教育部重点实验室,湖南长沙 410128

收稿日期:2011-01-10 修回日期:2011-03-03 出版日期:2011-06-15 发布日期:2019-09-06
通讯作者: larkin-liu@163.com
作者简介:李勤（1983— ）,男,湖南岳阳人,博士,主要从事茶树种质资源改良。
基金资助:
国家自然科学基金“茶氨酸基因合成的基因转录调控研究”（30871572）

Protein Extraction Method Suitable for Tea Plant

LI Qin^1,2, HUANG Jian-an^1,2, LIU Shuo-qian¹, LI Juan^1,2, LIU Zhong-hua^1,2*

1. National Research Center of Engineering Technology for Utilization of Functional Ingredients From Botanicals, Changsha 410128, China;
2. Key Lab of Tea Science, Ministry of Education, P.R. C., Changsha 410128, China

Received:2011-01-10 Revised:2011-03-03 Online:2011-06-15 Published:2019-09-06

摘要/Abstract

摘要： 为探索一种适用于茶树蛋白质双向电泳的样品制备方法,研究比较了TCA/丙酮沉淀法、改良的Tris-HCl抽提法,以及酚-甲醇/醋酸铵沉淀法3种植物蛋白质样品制备方法。结果表明,改良的Tris-HCl抽提法较适合于茶树蛋白质双向电泳的样品制备。该改良方法制备的茶树蛋白质样品经双向电泳分离,可获得1117±9.89个蛋白质点,其中大部分蛋白质的等电点集中在pI5~7之间,相对分子质量集中在15.00~95.00kD之间,背景清晰且蛋白质得率较高,能满足茶树蛋白质组学研究的需要。

关键词: 茶树, 双向电泳, 蛋白质样品制备, 蛋白质组学

Abstract: In order to develop an efficient protein extraction method suitable for two-dimensional electrophoresis from tea plant, three methods (TCA/acetone precipitation, improved Tris-HCl extraction and phenol-methanol/ammonium acetate precipitation) for total protein extraction in plant were compared. The result showed that the improved Tris-HCl extraction was the best method suitable for tea leaf protein sample preparation in these three techniques and the extracts were separated by two-dimensional electrophoresis. Consequently, 1117±9.89 protein spots were found and their isoelectric points were found to lie between pI5~7. The relative molecular weight of proteins was between 15.00~95.00kD. The improved Tris-HCl extraction could get the highest resolution of two dimensional electrophoresis and high yield of protein, and it could meet the requirement for the research on tea proteomecis.

Key words: Camellia sinensis (L.) O.Kuntze, two dimensional electrophoresis, protein sample preparation, proteomics

中图分类号:

S571.1
Q503

李勤, 黄建安, 刘硕谦, 李娟, 刘仲华. 茶树蛋白质双向电泳样品制备技术研究[J]. 茶叶科学, 2011, 31(3): 173-178. doi: 10.13305/j.cnki.jts.2011.03.014.

LI Qin, HUANG Jian-an, LIU Shuo-qian, LI Juan, LIU Zhong-hua. Protein Extraction Method Suitable for Tea Plant[J]. Journal of Tea Science, 2011, 31(3): 173-178. doi: 10.13305/j.cnki.jts.2011.03.014.

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茶树蛋白质双向电泳样品制备技术研究

Protein Extraction Method Suitable for Tea Plant

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