凹叶厚朴叶片蛋白提取及双向电泳体系的优化  

Total protein extraction and optimization of 2-DE system from the leaf proteome of Magnolia officinalis

作　　者：陈剑成 沈少炎 蔡月琴[1,4] 杨德明[1] 陈思凯[1] 万娟[1] 徐雯[1] 何天友[3] 郑郁善[1,3,2] 陈礼光 CHEN Jiancheng SHEN Shaoyan CAI Yueqin YA NG Deming CHEN Sikai WAN Juan XU Wen HE Tianyou ZHENG Yushan CHEN Liguang(College of Forestry, Fujian Agrieuhure and Forestry University,Fuzhou 350002,China College of Art College, Landscape Architecture, Fujian Agriculture and Forestry University,Fuzhou 350002, China Fujian GAP Engineering Research Center of Chinese Medicinal Crops, Fuzhou 350002,China College of Biological Sciences and Teehnology,Minnan Normal University,Zhangzhou 363000,China)

机构地区：[1]福建农林大学林学院,福建福州350002 [2]福建中药材GAP工程技术研究中心,福建福州350002 [3]福建农林大学艺术园林学院,福建福州350002 [4]福建闽南师范大学生物科学与技术学院,福建漳州363000

出　　处：《北方农业学报》2017年第2期11-16,共6页Journal of Northern Agriculture

基　　金：国家科技支撑计划项目(2011BAI01B06);福建省科技重点项目(2009Y0003)

摘　　要：为建立一套适于凹叶厚朴幼叶的双向电泳(2-DE)体系,以凹叶厚朴幼叶为材料,对蛋白提取方法、上样量、聚焦程序优化,试验结果表明,TCA/丙酮+Tris-酚法提取蛋白干粉最佳,具有产量高、污染低、背景浅特点;选用17 cm pH5~8NL IPG预制胶条、15μg上样量、聚焦程序Ⅱ的2-DE电泳为最适凹叶厚朴蛋白双向电泳体系。该体系为后续凹叶厚朴分子机理研究奠定基础。The purpose of this study is to establish an suitable two-dimensional electrophoresis （2-DE） for Magnolia officinalis leaf protein.The fresh leafs were chosen and grinded in the experiment of two-dimensional gel electrophoresis. The results showed TCA/acetone＋Tris saturated phenol method with clear background intensity, high resolution and purity in the content of separation protein. In consideration of the optimized system through selection of separating the proteins of leaves with 17 cm pHS-SNL IPG strips and IEF II and 2-DE program. It provided the reference for research progress in proteomics of species Magnolia officinalis.

关 键 词：凹叶厚朴 叶片 条件优化 蛋白质组 双向电泳(2-DE) 

分 类 号：S567.11[农业科学—中草药栽培]