摘要
对适用于秋茄(Kandelia candel)叶片蛋白质组学研究的双向电泳技术体系进行了优化。结果表明,采用酚抽提法提取的蛋白质浓度较低,约1.7μgμL-1,SDS-PAGE电泳后几乎无条带;而采用改良TCA-丙酮沉淀法可显著提高提取液中蛋白质的含量,可达12.4μgμL-1。秋茄叶片蛋白质主要分布在pH 4～7的范围内,最适于采用pH 3～10 NL的胶条,17 cm胶条的最适上样量为400μg,经银染后可得蛋白点约(564±34)个。用TCA-丙酮沉淀法提取秋茄叶片全蛋白质组,按优化的参数进行双向电泳分离,获得了高分辨率、高重复性的图谱,可用于秋茄蛋白质组学研究。
In order to study the pmteome of Kandelia candel leaves, a two-dimensional electmphoresis （2-DE） system was established and opthxized. Corrpared with the low protein content （1.7 μg ·L^-1） extracted by phenol （Phe）, the improved trichlomacetic acid/acetone precipitation （TCA） rrethod could obviously increase the extracting efficiency up to 12A μg ·L^-1. The protein spots ofKandelia candel leaves 1-minly distnbuted in the range ofpH 4- 7. The optimal loading quantity of samples were 400μg gel^-1 for a 17 cm IPG strip （pH 3-10 NL）, which can detect 564 ±34 protein spots after silver staining. Therefore, the high resolution and repeatable 2-DE nap could be obtained by using TCA extract nethod and the optimum system, which could widely use in proteome study of Kandelia candel.
出处
《热带亚热带植物学报》
CAS
CSCD
北大核心
2011年第6期519-523,共5页
Journal of Tropical and Subtropical Botany
基金
国家海洋公益性行业科研专项经费项目(200805072
201005012)
浙江省近岸水域生物资源开发与保护重点实验室开放课题基金项目(2010F30003)
浙江省科技厅创新团队建设与人才培养项目(2009F20009)
浙江省海洋与渔业项目“高纬度红树林生态恢复关键技术应用示范”资助
关键词
秋茄
蛋白质
提取
双向电泳(2-DE)
技术优化
Kandelia candel
Protein
Extraction
Two-dirrensional electrophoresis （2-DE）
Opt^tion
