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油菜根系蛋白双向电泳技术体系的建立

发表时间:2011-10-15  浏览量:1686  下载量:705
全部作者: 王志方,王振华,陈水森,石磊,徐芳森
作者单位: 华中农业大学作物遗传改良国家重点实验室,华中农业大学微量元素研究中心
摘 要: 通过对IPG胶条pH值范围、胶条长度、凝胶浓度以及等电聚焦条件、SDS-PAGE电压条件等技术参数的摸索优化,建立了适用于甘蓝型油菜根系蛋白质组学研究的双向凝胶电泳(two dimensional electrophoresis,2-DE)技术体系:采用18 cm、pH值为5~8的线性IPG胶条进行等电聚焦,SDS-PAGE分离采用质量分数为12%的聚丙烯酰胺凝胶,于50 V低压进样1 h后调整至120 V进行蛋白分离、染色后可获得较理想的蛋白质2-DE图谱。
关 键 词: 植物营养学;双向电泳;甘蓝型油菜;蛋白质组学;技术体系
Title: Establishment of a two-dimensional electrophoresis system for Brassica napus root protein
Author: WANG Zhifang, WANG Zhenhua, CHEN Shuisen, SHI Lei, XU Fangsen
Organization: National Key Laboratory of Crop Genetic Improvement, Microelement Research Center, Huazhong Agricultural University
Abstract: In this paper, optimized technical system of two-dimensional electrophoresis (2-DE) including the gel strength, pH range and length of IPG strip, technical parameters in IEF and SDS-PAGE was screened, and a 2-DE system was finally set up for proteome research of Brassica napus root, e.g. 18 cm IPG strips with liner pH 5-8 were adopted in IEF, 12% gel concentration was used in SDS-PAGE, the SDS-PAGE was performed in a low voltage of 50 V firstly to ensure protein sample getting into gels, then changed to 120 V for protein separation.
Key words: plant nutrition; two-dimensional electrophoresis; Brassica napus; proteomics; technical system
发表期数: 2011年10月第19期
引用格式: 王志方,王振华,陈水森,等. 油菜根系蛋白双向电泳技术体系的建立[J]. 中国科技论文在线精品论文,2011,4(19):1742-1748.
 
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