森林与环境学报2017，Vol.37Issue(1)：54-59,6.DOI:10.13324/j.cnki.jfcf.2017.01.009
福建柏叶片蛋白质双向电泳技术优化
Protocols optimization in two-dimensional electrophoresis of Fokienia hodginsii leaves total protein
摘要
Abstract
In order to study the most efficient 2-DE protocol of Fokienia hodginsii proteomics, three different methods including TCA-acetone, phenol extraction method and TCA-phenol extraction method were tested in the experiment. Besides, three different IPG strips of pH3~10, pH5~8, pH4~7 combined with three different focusing parameter which showed 20 000, 25 000, 30 000 Vh respectively were performed in IEF. Finally, different concentration separation gel (10%,12.5%)were performed in all SDS-PAGE trial. The results indicated that more protein spots were detected on 2-DE gels by phenol extraction method, IPG strips of pH4~7, 25 000 Vh focusing parameter, and 10% separate gel were the best settings of the 2-DE protocol. The experiment preliminary established the 2-DE protocols of F. hodginsii leaves which would be beneficial for the further study of proteinmics.关键词
双向电泳/福建柏/蛋白质组/等电聚焦/pHKey words
two-dimensional electrophoresis/Fokienia hodginsii/proteomics/isoelectric focusing/pH分类
农业科技引用本文复制引用
杨德明,张娅欣,沈少炎,荣俊冬,何天友,郑郁善..福建柏叶片蛋白质双向电泳技术优化[J].森林与环境学报,2017,37(1):54-59,6.基金项目
特色林木种质材料选育与高效培育关键技术研究( K81MLV09A)。 （ K81MLV09A）
