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园艺学报 ›› 2011, Vol. 38 ›› Issue (7): 1341-1348.

• 研究报告 • 上一篇    下一篇

‘秦冠’苹果MdWRKY基因亚细胞定位及原核表达

高 华1,2,樊红科1,党志国1,王 飞1,王雷存1,2,刘振中1,2,赵政阳1,2,*   

  1. (1西北农林科技大学园艺学院,陕西杨凌 712100;2陕西省苹果工程技术研究中心,陕西杨凌 712100)
  • 收稿日期:2011-04-07 修回日期:2011-06-08 出版日期:2011-07-25 发布日期:2011-07-25
  • 通讯作者: 赵政阳1,2,*

Studies on Subcellular Localization and Expression in E.coli of MdWRKY from‘Qinguan’Apple

GAO Hua1,2,FAN Hong-ke1,DANG Zhi-guo1,WANG Fei1,WANG Lei-cun1,2,LIU Zhen-zhong1,2,and ZHAO Zheng-yang1,2,*   

  1. (1College of Horticulture,Northwest A & F University,Yangling,Shaanxi 712100,China;2Apple E & T Research Centre of Shaanxi Province,Yangling,Shaanxi 712100,China)
  • Received:2011-04-07 Revised:2011-06-08 Online:2011-07-25 Published:2011-07-25
  • Contact: ZHAO Zheng-yang1,2,*

摘要: 以‘秦冠’苹果为试材,采用RT-PCR方法获得了一个WRKY基因,全长1 224 bp,推断其编码331个氨基酸,命名为MdWRKY。亚细胞定位分析MdWRKY蛋白分布在细胞核内,属于核蛋白。实时定量分析结果显示,MdWRKY基因受苹果斑点落叶病菌的诱导,表明其可能参与植物与病原菌的互作。随后进行原核表达分析,SDS/PAGE电泳结果表明表达蛋白与其蛋白大小一致。

关键词: 苹果, 转录因子, 克隆, 亚细胞定位, 表达

Abstract: A WRKY gene,designated as MdWRKY,was isolated from‘Qinguan’apple leaves using reverse transcription PCR approaches. MdWRKY was 1 224 bp and encoded a 331 amino-acid protein. Analysis of subcellular localization indicated that MdWRKY protein was targeted to the nucleus. Quantitative RT-PCR analysis showed that MdWRKY transcription was induced by Alternaria alternata f sp. mali indicating that MdWRKY gene may be involved in the interaction between plant and pathogen. Then expression in E. coli was conducted,SDS/PAGE result demonstrated that expression proteins consistent with the size of expected protein,which provided a foundation for further purifying and function study of MdWRKY.  

Key words: apple, transcription factors, clone, subcellular localization, expression

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