烟草叶片蛋白质组双向电泳实验体系的建立  

Establishment of Two-dimensional Electrophoresis Protocol for Tobacco Leaf Proteome

作　　者：王绍美[1] 罗成刚[1] 刘贯山[1] 冯全福[1] 苏振刚[1] 陈雅琼[1] 蒋彩虹[1] 孙玉合[1] 

机构地区：[1]烟草行业烟草遗传育种重点实验室,中国农业科学院烟草研究所,青岛266101

出　　处：《中国烟草科学》2012年第6期54-60,共7页Chinese Tobacco Science

基　　金：中国农业科学院中央级公益性科研院所基本科研业务费专项"干旱胁迫下烟草蛋白质组分的差异分析研究"(0032010037);中国农业科学院作物科学研究所中央级公益性科研院所基本科研业务费专项"黑胫病菌诱导的烟草蛋白质组分差异分析研究及差异蛋白质的鉴定分析研究"(201011);青岛市科技计划项目"应用蛋白质组学技术筛选与鉴定茄科作物抗黑胫病相关基因"[11-2-4-5-(12)-jch]

摘　　要：选用干旱敏感型栽培烟草品种中烟100旺长后期的叶片和感黑胫病栽培烟草品种小黄金1025苗期的叶片作为实验材料,总蛋白质提取分别采用稍加改进的TCA/丙酮法和由蛋白质分级提取、酚抽提及丙酮洗涤相结合的方法。对中烟100旺长后期烟草叶片采用的总蛋白质TCA/丙酮提取方法进行了优化、对其使用的IPG胶条pH范围做了初步筛选,并在此研究基础上,对采用第二种提取方法获得的小黄金1025苗期烟草叶片总蛋白质样品进行了不同上样量对双向电泳结果影响的比较实验。初步建立起适合烟草叶片蛋白质组研究的双向电泳实验体系。The plant materials used in these experiments were the flourishing late tobacco leaves of Zhongyan 100, a drought-sensitive variety, and the seedling leaves of Xiaohuangjin 1025, a variety susceptible for the tobacco black shank pathogen. Total proteins were extracted by trichloroacetic acid / acetone precipitation method, respectively, and protein fractionated extraction combined with phenol extraction and acetone washing method with some minor modifications. A trichloroacetic acid / acetone precipitation method, used for protein extraction from Zhongyan 100 leaves, was optimized. The immobilized pH gradient strips applied to Zhongyan 100 total proteins was selected preliminarily. On the basis of this study, we compared the effects of different protein quantities, from Xiaohuangjin 1025 total proteins extracted by the second protein extraction method, on 2-DE profile. 2-DE experiment protocol suitable for tobacco leaves proteome study was established preliminarily.

关 键 词：烟草 叶片 蛋白质组 双向电泳 

分 类 号：S572[农业科学—烟草工业]