巴西橡胶树树皮蛋白质组学分析体系的构建  

Preparation and MALDI-TOF MS analysis of bark proteins in Hevea brasiliensis

作　　者：袁坤[1] 王真辉[1] 丁璇[1,2] 杨礼富[1] 

机构地区：[1]中国热带农业科学院橡胶研究所,农业部橡胶树生物学重点开放实验室,海南儋州571737 [2]海南大学农学院,海南儋州571737

出　　处：《南京林业大学学报（自然科学版）》2009年第6期143-146,共4页Journal of Nanjing Forestry University：Natural Sciences Edition

基　　金：海南省自然科学基金资助项目(30819);中央级公益性科研院所基本科研业务费专项资助项目(XJSYWFZX2008-12)

摘　　要：巴西橡胶树（Hevea brasiliensis）是一种重要的产胶植物，其体内合成和贮存胶乳的地方是乳管系统，而乳管主要分布于树皮中。以10年生巴西橡胶树无性系“热研88—13”为研究对象，采用改进的三氯乙酸（TCA）一丙酮沉淀法提取树皮蛋白，使用17cm，pH4～7的IPG胶条进行双向电泳，得到良好的蛋白图谱。PDQuest软件（Bio—rad）分析银染后的凝胶，共得到约350个蛋白点。从这些蛋白点中随机选取10个蛋白点经基质辅助激光解析电离飞行时间质谱（MALDI—TOFMS）分析后，搜索NCBInr数据库对蛋白进行鉴定。结果显示，这些蛋白主要包括两种来源于巴西橡胶树中的重要蛋白，即橡胶小粒子蛋白（Small rubber particle protein）和橡胶树凝集因子（herein），此外还有一些其他功能和未知功能的蛋白。Hevea brasiliensis is an important plant of latex production. The place of latex synthesis and storage is the la- ticifer system, which mostly distributes in bark. In this study, the ten-year-old clone ＂Reyan 88 -13＂ was used as the experiment material. Bark proteins were extracted adopting improved TCA-acetone method. Using pH 4--7 IPG strip of 17cm to perform two-dimensional gel electropboresis （2-DE） , a good protein pattern was showed. PDQuest software （Bio-rad） analysis on the silver stained gel, about 350 protein spots were obtained. Ten of them were randomly selected and subjected to Matrix assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS） followed by searching NCBInr database to identify these proteins. The results showed that the proteins mainly included two impor- tant proteins in H. brasiliensis, namely small rubber particle protein and hevein. Some other function and unknown pro- teins were also found. It has important significance for biology research that the establishment of 2-DE system and primary MS analysis of bark proteins in H. brasiliensis.

关 键 词：巴西橡胶树 树皮 蛋白质组学 双向电泳 MALDI—TOF MS 

分 类 号：S722[农业科学—林木遗传育种]