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

作　　者：杨德明[1,2] 张娅欣 沈少炎 荣俊冬[1] 何天友[3] 郑郁善[1] 

机构地区：[1]福建农林大学林学院,福建福州350002 [2]福建农林大学工业原料研究所,福建福州350002 [3]福建农林大学艺术园林学院,福建福州350002

出　　处：《森林与环境学报》2017年第1期54-59,共6页Journal of Forest and Environment

基　　金：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)

摘　　要：蛋白质双向电泳是蛋白质组学研究方法之一,但蛋白质双向电泳技术体系却因物种不同而有所差异,因而蛋白质双向电泳体系的建立是进行蛋白质分离、鉴定的首要步骤。本文对福建柏叶片蛋白质提取方法、IPG胶条、等电聚焦参数设定、丙烯酰胺分离胶浓度等因子进行单因素分析,对福建柏双向电泳技术进行优化。结果表明,采用三氯乙酸-丙酮提取福建柏叶片的蛋白点多且图像清晰,p H4~7的IPG胶条最适合福建柏叶片蛋白的分离,等电聚焦参数设定为20 000 Vh,浓度为10%的分离胶进行福建柏双向电泳最好。本研究初步建立了福建柏叶片蛋白质双向电泳体系,为深入研究福建柏蛋白质组学提供依据。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 p H3 - 10,p H5 - 8,p H4 - 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 p H4 -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 PH 

分 类 号：Q946.1[生物学—植物学]