首页 | 本刊简介 | 编委会 | 投稿征稿 | 期刊订阅 | 下载中心 | English
 
   
  论文快速检索

请输入关键词

 

 

   
  信息公告
海外撷英栏目开通,欢迎投稿

热带亚热带植物学报网络开通

访问总次数
13812022
今日访问
3444
当前在线
0人
 
引用本文:
【打印本页】   【HTML】   【下载PDF全文】   查看/发表评论  【EndNote】   【RefMan】   【BibTex】
←前一篇|后一篇→ 过刊浏览    高级检索
本文已被:浏览 1034次   下载 860 本文二维码信息
码上扫一扫!
分享到: 微信 更多
菠菜乙醇酸氧化酶同工酶GO Ⅰ的纯化和特性
徐杰, 吴燕燕
作者单位
徐杰, 吴燕燕  
摘要:  从菠菜绿叶中获得SDS-PAGE为40 000±2 000Mr单带的乙醇酸氧化酶同工酶GO I。其GO比活为8.4 U min-1mg-1 protein,经ND-PAGE后显示Mr为470 000。用IEF测得其等电位为7.4。用氧电极法证实该GO I能同时催化乙醇酸和乙醛酸的氧化,制备GO I的抗体并对菠菜绿叶粗蛋白作免疫双扩散,有免疫沉淀线,经蛋白A-Sepharose CL-4B柱吸附法和抗原亲和吸附法,得到单特异性GO I抗体,用该GO I抗体对菠 菜绿叶粗蛋白作SDS-PAGE Western blot后只有40 000±2 000-一条染色带,作4%-20%PAGE Western lot后有多条 色带,Mr分别大约为470 000,280 000和80 000,表明菠菜绿叶中的GO I有多个聚合态。
关键词:  乙醇酸氧化酶同工酶  菠菜  催化特性  等电点  分子量  聚合态
DOI:10.3969/j.issn.1005-3395.2001.1.003
分类号:
基金项目:
PURIFICATION AND PROPERTIES OF GLYCOLATE OXIDASE ISOZYME GO I FROM SPINACH LEAVES
XU Jie, WU Yan-yan
Abstract:  By using ammonium sulfate fractionation, Sephadex G-25, DEAE-Cellulose, and Sepharose-6B chromatographies, glycolate oxidase isozyme GO I was purified from spinach green leaves which showed only one band with molecular weight (Mr) of 40 000± 2 000 in SDS-PAGE. The specific activity of purified GO I was 8.4 U min-1mg-1 protein. GO I showed its Mr of 470 000 in 4%-20% ND-PAGE and could catalyze both oxidations of glycolate and glyoxylate simultaneously. The isoelectric point (PI) of GO I was about PH 7.4. There was a immunity precipitate line determined by immunity double-difusion reaction between the antibody raised against GO I and crude protein from spinach green leaves.The specitic G0 I antibody was purified by protein A-Sepharose CL-4B column and specific antigen affinity elution.Only the 40 000± 2 000 band could immunity cross-react with Go I antibody,and different Go I polvmers of 470 000,280 000,and 80000 were found when crude protein were conduced to SDS-PAGE Ⅵ stern blot or ND-PAGE Western blot using this speciftc GO I antibody.
Key words:  Glycolate oxidase isozyme  Spinach  Catalyzed property  Isoelectric point  Molecular weight  Polymeric form

Copyright 2007 版权所有 中国科学院华南植物园《热带亚热带植物学报》编辑部
地址: 广州市天河区兴科路723号 中国科学院华南植物园 《热带亚热带植物学报》编辑部 邮编:510650 电话: 86 (0)20-3725 2514  传真:86 (0)20-3725 2642 E-mail: jtsb@scib.ac.cn
本系统由北京勤云科技发展有限公司设计
京ICP备09084417号-2