茶树雌蕊总蛋白质双向电泳分离体系的建立

doi:10.13305/j.cnki.jts.2013.05.012

茶叶科学 ›› 2013, Vol. 33 ›› Issue (5): 420-428.doi: 10.13305/j.cnki.jts.2013.05.012

茶树雌蕊总蛋白质双向电泳分离体系的建立

任燕^1,2, 陈暄¹, 房婉萍¹, 李垚¹, 黎星辉^1,2,*

1. 南京农业大学园艺学院,江苏南京 210095;
2. 苏州市现代生态茶业工程技术研究中心,江苏苏州215128

收稿日期:2013-04-22 修回日期:2013-07-27 出版日期:2013-10-30 发布日期:2019-09-04
通讯作者: *lxh@njau.edu.cn
作者简介:任燕（1989— ）,女,四川广元人,硕士研究生,主要从事茶树育种与栽培研究。
基金资助:
国家自然科学基金（30972403）、江苏高校优势学科建设工程资助项目（XDYY2011）

Establishment of Two-dimensional Electrophoresis System for Pistil Proteome Analysis in Tea Plant

REN Yan^1,2, CHEN Xuan¹, FANG Wan-ping¹, LI Yao¹, LI Xing-hui^1,2,*

1. College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;
2. Suzhou Engineering Research Center for Modern Ecological Tea Industry, Suzhou 215128, China

Received:2013-04-22 Revised:2013-07-27 Online:2013-10-30 Published:2019-09-04

摘要/Abstract

摘要： 为建立一套适合茶树雌蕊总蛋白质分离的双向电泳体系,对茶树雌蕊总蛋白的提取方法、SDS-PAGE凝胶浓度、聚焦条件和IPG胶条pH范围等进行了比较优化。结果表明,TCA/丙酮-clean up kit法提取总蛋白,用17βcm pH 5~8 IPG胶条,13% SDS-聚丙烯酰胺凝胶进行分离,用考马斯亮蓝R250染色,可以获得清晰、分辨率高、重复性好的双向电泳图谱,共检测到约1β200个蛋白点,主要分布在pH 5~8,相对分子量14~117βkD范围内,碱性蛋白得到有效分离,可以满足茶树雌蕊的蛋白组学分析和研究。

关键词: 茶树, 雌蕊, 蛋白质提取, 双向电泳

Abstract: In order to establish a proper two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of tea plant pistils, several protein extraction methods, density of SDS-PAGE gel, IEF procedures and pH gradient of IPG strip were compared. The results showed the optimized system includes: total proteins of sample extracted using TCA/acetone method followed with clean up, separating the proteins with 17βcm pH 5~8 IPG strips, 13% SDS-PAGE, and CBB R-250 stain method. About 1β200 spots can be detected, and these proteins mainly distributed in the pH 5~8, MW 14~117βkD range. With good separation of the basic protein, this will be helpful to proteomic research in tea plant.

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

中图分类号:

S571.1
Q51

任燕, 陈暄, 房婉萍, 李垚, 黎星辉. 茶树雌蕊总蛋白质双向电泳分离体系的建立[J]. 茶叶科学, 2013, 33(5): 420-428. doi: 10.13305/j.cnki.jts.2013.05.012.

REN Yan, CHEN Xuan, FANG Wan-ping, LI Yao, LI Xing-hui. Establishment of Two-dimensional Electrophoresis System for Pistil Proteome Analysis in Tea Plant[J]. Journal of Tea Science, 2013, 33(5): 420-428. doi: 10.13305/j.cnki.jts.2013.05.012.

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茶树雌蕊总蛋白质双向电泳分离体系的建立

Establishment of Two-dimensional Electrophoresis System for Pistil Proteome Analysis in Tea Plant

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