【作者】 陈富成； 祁建民； 徐建堂； 陈涛； 陶爱芬； 林培清； 陈美霞； 李华丽；

【Author】 Chen Fu-cheng,Qi Jian-min~*,Xu Jian-tang,Chen Tao,Tao Ai-fen,LIN Pei-qing,Chen Mei-xia,Li Hua-li (Key Laboratory of Ministry of Education for Genetics,Breeding and Multiple Utilization of Crops.Fujian Agriculture and Forestry University,Fuzhou,350002)

【机构】 福建农林大学作物遗传育种与综合利用教育部重点实验室；

【摘要】 蛋白质提取方法和电泳条件是蛋白质组学研究中双向电泳的关键。本文初步建立起一套适合黄麻功能叶总蛋白双向电泳(2-DE)分析的高效提取方法及电泳条件,采用双向电泳技术,分析比较了TCA丙酮法、Tris-base/丙酮法、Tris-HCl法三种提取方法,并对上样量和样品制备方法等进行了优化。结果表明:TCA丙酮法提取蛋白质产量高,达79.0mg/g,比Tris-base/丙酮法和Tris-HCl的产量分别高出44.2%,114.1%。该法得到的2-DE图谱背景清晰,蛋白点数最多,约602个,较其它两种方法更适合作为黄麻功能叶总蛋白双向电泳分析的提取方法;350ug为优化后考蓝时的蛋白最佳上样量。这为黄麻蛋白质组学后续研究奠定了基础。更多还原

【Abstract】 The protein extraction methods and electrophoresis conditions are the key of two -dimensional electrophoresis in proteomics.In this text,in order to set up a suitable method for jute leaf total protein extraction and two -dimensional electrophoresis condition,we used the two -dimensional electrophoresis to draw a comparison between TCA acetone method,Tris-base acetone method and Tris-HCl method.We also optimized the sample loading quantity and sample preparation approach.The results showed:the protein productivity was the highest reaching 79.0mg/g by TCA acetone method,which the productivity is higher than the two other methods and the values are44.2%and 114.1%respectively.Besides,the 2-DE image background was the clearest and the protein spots are the most reaching aboat 602 by TCA acetone method,it was much more suitable as extraction method for jute leaf protein two-dimensional electrophoresis.The most suitable sample loading quantity was 350 ug in coomassie brilliant blue,which laid the foundation for further study of jute proteomics.更多还原