文献类型：期刊文献

中文题名：青杨双向电泳实验体系的建立

英文题名：Populus leafy proteomics with NaCl stress using two-dimensional electrophoresis analysis

作者：陈国良[1,2] 蒋晶[1] 乔桂荣[1] 金梁[2] 孙月华[2] 杨晔[1] 周婧[1] 卓仁英[1]

第一作者：陈国良

机构：[1]中国林业科学研究院亚热带林业研究所;[2]兰州大学草地农业科技学院

年份：2009

卷号：26

期号：5

起止页码：644-651

中文期刊名：浙江林学院学报

外文期刊名：Journal of Zhejiang Forestry College

收录：北大核心:【北大核心2008】；CSCD:【CSCD_E2011_2012】；

基金：非盈利性科研院所基金资助项目(RISF060702)

语种：中文

中文关键词：植物学;氯化钠胁迫;蛋白质组学;双向电泳;青杨

外文关键词：botany; NaCl stress; proteomics; two-dimensional electrophoresis （2-DE）; Populus cathayana;

分类号：S718.4

摘要：用改良丙酮沉淀法、三氯乙酸(TCA)-丙酮沉淀法和酚抽法抽提青杨Populuscathayana的蛋白质,分别获得了437,603和721个蛋白斑点,以酚抽法提取的分离纯化方法效果最佳,不仅能很好地提取蛋白,而且能有效去除样品中的盐分。通过比较,分析了双向电泳不同条件对电泳结果的影响,确立了300μg.IPG胶条-1的最佳上样量、90000V.h的聚焦时间以及20min平衡时间的双向电泳条件。研究还比较了氯化钠胁迫前后双向电泳图谱差异,发现有46个蛋白点存在差异,其中10个下调,36个上调(4个新诱导表达)。
Salt stress is one of the major abiotic stressed over the world. Most of the researchs were foucsed on improve the ability of plant species living in salt soil. Two-dimesional gel electrophoresis （2-DE）is one of the most useful methods in proteins research. To determine total protein extraction efficiency from Populus cathayana with two-dimensional electrophoresis （2-DE） analysis,three protein extraction methods （phenol extraction,improved acetone precipitation method,and TCA-acetone precipitation method） were compared.Electrophoresis conditions and leafy proteomics of P. cathayana with salt stress were also analyzed. Results showed that the three protein extraction methods generated 437, 603, and 721 protein spots with the phenol extraction method more effectively removing salt from the protein sample, generating more protein spots, and giving clearer background for the 2-DE map. The results showed that condition of 300 μg per IPG gel, 20 min equilibrium time, 90 000 V· h IEF time was the suitable treatment. Also, the leafy proteomies analysis revealed that 10 protein spots was down-regulators, 32 protein spots was up-regulators, and 4 new protein spots was induced by salt stress. These proteins may be important response to salt stress.