大白菜根系蛋白质双向电泳体系优化  

Optimizing Two-Dimensional Gel Electrophoresis System for Root Proteins of Chinese Cabbage(Brassica rapa L. ssp. Pekinensis)

作　　者：赵玲玲[1] 毕青[1] 周雪[1] 冀瑞琴[1] 冯辉[1] 

机构地区：[1]沈阳农业大学园艺学院、辽宁省十字花科蔬菜遗传育种重点实验室,沈阳110866

出　　处：《分子植物育种》2015年第1期178-183,共6页Molecular Plant Breeding

基　　金：国家自然科学基金(312721-57);辽宁省自然科学基金(2013020071)共同资助

摘　　要：双向电泳技术是蛋白质实验室最常用的手段之一,已被广泛应用于蛋白质组学研究中。为建立适合大白菜根部蛋白质组分析的双向电泳体系,得到比较完美的电泳图谱,本研究着重从等电聚焦参数设置、IPG胶条p H范围、上样量,胶体浓度条件进行了优化。结果表明：选择17 cm p H 4~p H 7的IPG胶条,上样量为1 200μg,聚焦时间为85 000 Vh,胶体浓度为12.5%进行双向电泳,考马斯亮蓝染色可得到背景清晰、重复性好、蛋白点分辨率较高的大白菜根部双向电泳图谱。研究结果有助于开展大白菜根部病害及根部相关特性的蛋白质组学研究。As one of the most popular protein approaches in laboratory, two-dimensional electrophoresis （2-DE） had been widely applied to proteomic research. In order to establish an idea 2-DE system for proteomic analysis of Chinese cabbage root and to obtain perfect 2-DE profiles, 2-DE system for Chinese cabbage root was optimized based on some key factors, such as the isoelectric focusing time, the immobilized pH gradient （IPG）, sample load- ing amount and gel concentrations. In this research, the improved 2-DE system for the root of Chinese cabbage were come out as follows： IPG strips of 17 cm in length with the pH ranges from 4 to 7, 1 200 μg loading amount, 85 000 Vh of IEF time, and 12.5% gel concentration. Following this improved system, a perfect 2-DE profile of protein spots with clear background, good reproducibility and high resolution was stable generated by staining of the dye coomassie brilliant blue. This research might facilitate to carrying out proteomic studies related to root disease and root characteristics of Chinese cabbage.

关 键 词：大白菜 蛋白质 双向电泳 优化 

分 类 号：S634.1[农业科学—蔬菜学]