适用于花叶唐竹叶片总蛋白的双向电泳体系的建立  

Establishment of Two-dimensional Electrophoresis System Suitable for Total Protein of Sinobambusa tootsik Leaves

作　　者：陈凌艳[1] 赖金莉 张力 赵婷 何天友[1] 张赛 郑郁善[1,2] Chen Lingyan;Lai Jinli;Zhang Li;Zhao Ting;He Tianyou;Zhang Sai;Zheng Yushan(College of Landscape Architecture. Fujian Agriculture and Forestry University, Fuzhou, 350002;College of Forestry, Fujian Agriculture andForestry University, Fuzhou, 350002)

机构地区：[1]福建农林大学园林学院,福州350002 [2]福建农林大学林学院,福州350002

出　　处：《分子植物育种》2019年第7期2303-2311,共9页Molecular Plant Breeding

基　　金：福建省教育厅中青年教师教育科学研究项目(JAT170197);福建省科技重大专项(2013NZ0001);福建省科技重大项目(2011N5002)共同资助

摘　　要：本研究以花叶唐竹二年生苗木的健康叶片为材料,采用3种不同的蛋白质提取方法(酚法, TCA-丙酮法, TCA-丙酮/酚法)、不同的上样量、不同p H值IPG胶条及不同等电聚焦方案等因素进行筛选和优化,以期获得最佳的花叶唐竹叶片总蛋白的提取方法,并建立适用、高效的花叶唐竹蛋白质双向凝胶电泳技术体系。结果表明,选择TCA-丙酮/酚法的蛋白得率高、纯度高,选择17 cm pH 5～8的线性IPG胶条,450μg的上样量以及60 000 Vhs的等电聚焦强度进行双向电泳,共检测到726个蛋白点,整体分布均匀,分离效果较好。本研究初步建立了花叶唐竹蛋白质双向电泳体系,有助于后续研究叶片呈色机理提供技术帮助。In order to obtain the best extraction method of total protein of Sinobambusa tootsik and establish a suitable and efficient two-dimensional electrophoresis(2-DE) system for Sinobambusa tootsik proteomics analysis,the healthy leaves of biennial seedlings of Sinobambusa tootsik were used as materials in this study. different sample amount, different p H value IPG strips and different electrofocusing schemes were selected and optimized.Three protein extraction methods(phenol method, TCA-acetone precipitation method, and the TCA-acetone/phenol method), the pH range of IPG strips and isoelectric focusing programs were used for screening and optimization.The results showed that the TCA-acetone/phenol method selected could get a high purity and a high extraction rate. The effect of two-dimensional electrophoresis was excellent when conducted with immobilized 17 cm pH 5~8 IPG strips and 450 μg sample that 726 protein spots were detected, resulting in good separating effect and well distribution. This experiment preliminary established the 2-DE protocol of S. tootsik f. luteoloalbostriata leaves which would be beneficial for the further study of variegated leaves development mechanism.

关 键 词：花叶唐竹 双向电泳 蛋白组学 等电聚焦 

分 类 号：S795[农业科学—林木遗传育种]