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植物研究 ›› 2009, Vol. 29 ›› Issue (6): 692-695.doi: 10.7525/j.issn.1673-5102.2009.06.009

• 论文 • 上一篇    下一篇

高山离子芥CbMAPK3基因克隆与原核表达

张腾国1,3;刘玉冰2;孙坤1;杨宁1;安黎哲3*   

  1. (1.西北师范大学生命科学院,兰州 730070) (2.中国科学院寒区旱区环境与工程研究所,兰州 730000) (3.兰州大学生命科学院,兰州 730000)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-11-20 发布日期:2009-11-20
  • 通讯作者: 安黎哲
  • 基金资助:
     

Cloning and Prokaryotic Expression of Chorispora bungeana CbMAPK3 Gene

ZHANG Teng-Guo;LIU Yu-Bing;SUN Kun;YANG Ning;AN Li-Zhe*   

  1. (1.School of life science,Northwest Normal University,Lanzhou 730070) (2.Cold and Arid Regions Environmental and Engineering Institute,Chinese Academy of Sciences,Lanzhou 730000) (3.College of Life Science,Lanzhou University,Lanzhou 730070)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-11-20 Published:2009-11-20
  • Contact: AN Li-Zhe
  • Supported by:
     

摘要: 促分裂原活化蛋白激酶(MAPK)在信号传导过程中发挥着重要的作用。以高山离子芥叶片为材料,利用RT-PCR法克隆到全长CbMAPK3基因cDNA,将其与大肠杆菌表达载体pET-30a连接,构建原核表达载体pET-30a-CbMAPK3,并转化大肠杆菌BL21,经IPTG诱导表达后,SDS-PAGE以及Western blot检测结果表明该基因表达了1个约46kD的蛋白,为进一步研究目的蛋白的结构和功能提供了实验基础。

关键词: 高山离子芥, CbMAPK3基因, 原核表达

Abstract: Mitogen-activated protein kinase (MAPK) plays a central role in transfer information from diverse receptors/sensors to a wide range of cellular responses in plants. CbMAPK3 cDNA was amplified by RT-PCR from Chorispora bungeana leaf, then was cloned into pET-30a vector to construct recombinant prokaryotic expression vector pET-30a-CbMAPK3. The pET-30a-CbMAPK3 was transformed to E.coli strain of BL21. After induced by IPTG, a 46 kD recombinant protein was expressed and separated by SDS-PAGE electrophoresis and detected by western blot. The research provides a base for further studying on the protein structure and function of CbMAPK3.

Key words: Chorispora bungeana, CbMAPK3 gene, prokaryotic expression

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