首页 | 本学科首页   官方微博 | 高级检索  
   检索      

草木樨状黄芪变异系中甲硫氨酸代谢相关cDNA的分子克隆及其在大肠杆菌中的表达
引用本文:刘永军,景建洲,贾敬芬,李振勇.草木樨状黄芪变异系中甲硫氨酸代谢相关cDNA的分子克隆及其在大肠杆菌中的表达[J].西北植物学报,2004,24(1):1-5.
作者姓名:刘永军  景建洲  贾敬芬  李振勇
作者单位:1. 西北大学,生命科学学院,陕西省生物技术重点实验室,西安,710069
2. 华美生物工程公司博士后工作站,河南洛阳,471003;清华大学,生物技术系,北京,100063
3. 华美生物工程公司博士后工作站,河南洛阳,471003
基金项目:陕西省教育厅重点科研项目 02JS06
摘    要:利用mRNA差异显示技术(differential display)从草木樨状黄芪耐甲硫氨酸变异系中分离到一差异cDNA片段,命名为tm03(360bp)。mRNA杂交结果显示,该片段只在变异苗中表达,初步确定其与甲硫氨酸代谢相关。测序后在Genebank搜寻,未发现有同源序列。进一步分析表明,该cDNA克隆可编码完整的蛋白。在该基因片段两端分别合成含HindⅡ和BamH Ⅰ酶切位点的引物,用PCR扩增后亚克隆到pRSET A表达载体,转化宿主菌JMl09(DE3),经IPTG诱导表达了N′端含6个组氨酸的蛋白质,分子量约为14kD,为进一步研究该蛋白的结构和生物学功能奠定基础。

关 键 词:草木樨状黄芪变异系  甲硫氨酸代谢  克隆  表达
文章编号:1000-4025(2004)01-0001-05
修稿时间:2003年4月21日

Molecular cloning of methionine-metaboly correlated cDNA in Astragalus melilotoides Pall.variant line and its expression in E.coli
LIU Yong-jun,JING Jian-zhou.Molecular cloning of methionine-metaboly correlated cDNA in Astragalus melilotoides Pall.variant line and its expression in E.coli[J].Acta Botanica Boreali-Occidentalia Sinica,2004,24(1):1-5.
Authors:LIU Yong-jun  JING Jian-zhou
Institution:LIU Yong-jun~1,JING Jian-zhou~
Abstract:Differential cDNA fragment which was named tm03 (360 bp) was isolated from Astragalus melilotoides Pall. methionine-tolerant variant line by mRNA differential display analysis.After detected by mRNA dot hybridization,tm03 expressed only in the variant line,and tm03 was primarily confirmed as the methionine-metaboly correlated cDNA.tm03 was cloned and sequenced.After searching in GenBank,there were no similar sequences reported.Further research observed that the cDNA can code a complete protein.To construct high level expression plasmid,tm03 was ligated with the expression vector pRSET A,then transformed into JM109 (DE3) E.coli strain,and induced with IPTG.SDS-PAGE confirmed that protein of 6His-recombinant tm03 was expressed efficiently in inclusion bodies in E.coli,and the molecular weight was about 14 kD.It would be helpful to further study the structure of tm03 protein and its biological function.
Keywords:Astragalus melilotoides Pall  variant line  methionine-metaboly  clone  expression
本文献已被 CNKI 维普 万方数据 等数据库收录!
点击此处可从《西北植物学报》浏览原始摘要信息
点击此处可从《西北植物学报》下载免费的PDF全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号