一种适用于胡杨叶片非变性蛋白质提取的方法  

A Novel Method for Nondenaturing Protein Extraction from Populus Euphratica Leaves

作　　者：韩航 马燕芳 李征珍[1] 兰玉婷 石莎[1] 冯金朝[1] 王晓东 HAN Hang;MA Yan- fang;LI Zheng-zhen;LAN Yu-ting;SHI Sha;FENG Jin-chao;WANG Xiao-dong(College of Life and Environmental Sciences1,Centre for Imaging ＆ Systems Biology;Minzu University of China,Beijing 100081,China)

机构地区：[1]中央民族大学生命与环境科学学院,北京100081 [2]生物成像与系统生物学研究中心,北京100081

出　　处：《科学技术与工程》2018年第21期36-41,共6页Science Technology and Engineering

基　　金：国家自然科学基金(31770384,31570407)、中央民族大学青年学术团队引领计划项目(2017MDYL33)和中央民族大学本科创新项目基金(GCCX2017110019)资助

摘　　要：为探索适用于胡杨叶片非变性蛋白质的提取方法,以采自内蒙古额济纳旗天然胡杨林中胡杨成熟叶片为材料,采用咪唑法、Bis-Tris法、Tris-HCl法和新构建的Tris-SSAD法4种非变性蛋白质提取策略,分别用于提取胡杨叶片蛋白质样品;随后进行第一维非变性聚丙烯酰胺凝胶电泳(1^(st)-DE:Native-PAGE)。相比之下,只有Tris-SSAD法获得了清晰、条带数目较多的Native-PAGE图谱,且结合第二维变性凝胶电泳(2^(nd)-DE:SDS-PAGE)可成功分离出较多蛋白质复合体亚基或相互作用蛋白质分子,证明了Tris-SSAD法非变性温和提取特性。研究建立并优化了适用于胡杨叶片的非变性蛋白质提取方法,为后续进一步分析蛋白质与蛋白质相互作用及蛋白质复合体功能提供了实验基础。Aims to establisii a nondenaturing protein extracted metiiod to study protein complexes and interacting proteins from leaves of Populus euphratia, four approaches, including Imidazole, Bis-Tris,Tris-HCI,and new method of Tris-SSAD （Tris based Sucrose,Sodium chloride, Ascorbate,and Digitonin solution ）,were applied in the nondenaturing extraction of leaf proteins. Based on the first-dimensional non-denaturing gel electrophoresis （1st- DE ： Native- PAGE）,it was found that the highest quality protein bands on CBB stained gel were only observed in the protein extracted by the method of Tris- SSAD. Further optimized processing was also performed to obtain more perfect results. Meanwhile, the nondenatuing feature of Tris-SSAD were also successfully proven by the tandem electrophoresis,composed of nondenaturing 1st-DE （ Native-PAGE） and denaturing 2nd-DE （SDS-PAGE ）,and a lot of protein complexes and/or interactive proteins were found in these electrophoresis gels. Finally,a suitable for the nondenaturing protein extraction from adverse-resistant plant leaves was achieved.

关 键 词：胡杨 非变性蛋白质 非变性凝胶电泳 蛋白质复合体 相互作用蛋白质 

分 类 号：Q946.33[生物学—植物学]