大头典竹叶片蛋白质组学双向电泳技术体系的优化  

Optimization of Two-dimensional Polyaneylamide Gel Electrophoresis for Proteome of D.beecheyana var. pubescens

作　　者：陈思凯[1] 瞿印权 杨德明[1] 陈剑成 宋鲲鹏[1] 刘家琪[1] 何天友[2] 荣俊冬[1] 郑郁善[1,2] 陈礼光[1] Chen Sikai(College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350002, China)

机构地区：[1]福建农林大学林学院,福建福州350002 [2]福建农林大学艺术学院园林学院,福建福州350002

出　　处：《安徽农学通报》2017年第5期41-44,80,共5页Anhui Agricultural Science Bulletin

基　　金：福建省农业科技重大专项"特色林木种质材料选育与高效培育关键技术研究"(2013NZ0001);福建省科技重大项目"中小型竹资源优良品种选择和定向培育关键技术研究"

摘　　要：目的:为了获得清晰的蛋白凝胶图谱,建立适用于大头典竹蛋白组学双向电泳的优化体系。方法:对大头典竹蛋白提取方法、胶条的水化运行方式、等电聚焦(IEF)程序的设置、蛋白上样量等步骤进行优化。结果:大头典竹的蛋白斑点主要分布在pH5~8的范围;三氯乙酸-丙酮法与酚抽结合法更适合大头典竹蛋白的提取;IPG胶条胶面向上、被动水化方式、蛋白上样量为50μL,效果更好。结论:通过建立双向电泳(2-DE)的优化体系,获得的蛋白点均匀、清晰、横纵条纹少的凝胶图谱,提高了分辨率,为大头典竹差异蛋白组学研究奠定了基础。[ Objective ] In order to obtain a clear protein gel electrophoresis, the optimal two-dimensional electrophoresis system suitable for D.beecheyana var. pubescens was established. [Methods]The total protein extraction method, hydration operation mode of strip, program arguments setting of isoelectric focusing and protein amount were optimized to establish an optimal two-dimensional electrophoresis system suitable for D.beecheyana var.pubescens. [Results] Bamboo protein spots were mainly distributed in the scope of the pH5-8. Using trichloroacetic acid - acetone method combined with phenol extraction method is more suitable for the extraction of the D.beecheyana var. pubescens protein. Placing the surface of strip up-side down, passive hydration mode, 50μL amount of protein is benefit to obtain optimal effect of gel electrophoresis. [Conclusion] Through the establishment of two-dimensional electrophoresis （2 DE） the optimization of the system, obtaining protein point uniform, clear, map of transverse and less longitudinal stripes gel electrophoresis and improving the resolution, provided the basis for further studies of the proteomics for D.beecheyana var.pubescens.

关 键 词：大头典竹 双向电泳 蛋白组学 

分 类 号：Q946-33[生物学—植物学]