| 不同提取方法及上样量对牡丹试管苗茎基部蛋白双向电泳的影响 |
| |
| 引用本文： | 王海云,丁义,符真珠,何松林.不同提取方法及上样量对牡丹试管苗茎基部蛋白双向电泳的影响[J].广西农业生物科学,2010(1):91-96. |
| |
| 作者姓名： | 王海云 丁义 符真珠 何松林 |
| |
| 作者单位： | 河南农业大学林学院; |
| |
| 基金项目： | 河南省高校创新人才培养工程资助 |
| |
| 摘 要： | 为建立一套适合于牡丹试管苗茎基部蛋白的双向电泳技术,以便更好地利用蛋白质组技术研究牡丹试管苗不定根的发生机理,本研究比较了三种不同蛋白质提取方法对双向电泳结果的影响,并在蛋白质上样量方面进行了比较。结果表明,乙酸铵/甲醇酚提取法所得2-DE图谱的蛋白点很少,仅检测到45个,且较模糊,有明显的拖尾现象,分辨率很低;乙醇/乙醚丙酮法所得的蛋白点也较少（101个）,较模糊,且横竖纹干扰较大;三氯乙酸/丙酮法所得蛋白点数较多,可检测到434个清晰的蛋白点,且形状规则,重复性好,适合后续分析,操作也较为简便。用三氯乙酸/丙酮法提取蛋白,采用800μg、1000μg和1200μg三个不同的上样量进行双向电泳,在上样量为1200μg时（IPGpH3～10,24cm）,蛋白质在12%SDS-PAGE胶上得到了较好的分离,在2-DE图谱上可分辨出562个蛋白点。因此,三氯乙酸/丙酮法是较适合于牡丹试管苗茎基部蛋白质提取的方法,1200μg是较为合适的上样量。
|
| 关 键 词： | 牡丹 双向电泳 提取方法 上样量 |
Effects of Different Protein Extraction Methods and Loading Amount on 2-DE Profile for Basal Part of Mudan in vitro Stem |
| |
| Wang Haiyun Ding Yi Fu Zhenzhu He Songlin College of Forestry,He'nan Agricultural University,Zhengzhou.Effects of Different Protein Extraction Methods and Loading Amount on 2-DE Profile for Basal Part of Mudan in vitro Stem[J].Journal of Guangxi Agricultural and Biological Science,2010(1):91-96. |
| |
| Authors: | Wang Haiyun Ding Yi Fu Zhenzhu He Songlin College of Forestry He'nan Agricultural University Zhengzhou |
| |
| Institution: | Wang Haiyun Ding Yi Fu Zhenzhu He Songlin College of Forestry,He'nan Agricultural University,Zhengzhou,450002 |
| |
| Abstract: | In this paper,we compared the effects of different protein extraction methods and loading amount on 2-DE profile in order to establish a suitable and efficient 2-DE technique for proteome of basal part of stem of mudan in vitro and facilitate rooting mechanism research by using proteome approach. The results showed that samples prepared by ammonium acetate/methanol-phenol extraction only got 45 protien spots on the 2-DE profile,and the spots were indistinct,had obvious streak appearance and low resolution. ... |
| |
| Keywords: | Mudan （Paeonia suffruticosa） Two-dimensional electrophoresis （2-DE） Methods of protein extrac-tion Loading quantity of sample |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
