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园艺学报 ›› 2009, Vol. 36 ›› Issue (11): 1635-1642.

• 蔬菜 • 上一篇    下一篇

拟南芥抗病基因FLS2LysM的亚克隆及其在番茄中的表达

陈武1;杨玉婷1;李颖2;黄三文2;黎定军1*   

  1. (1 湖南农业大学生物安全科技学院, 长沙410128; 2 中国农业科学院蔬菜花卉研究所, 北京100081)
  • 收稿日期:2009-04-27 修回日期:2009-06-05 出版日期:2009-11-25 发布日期:2009-11-25
  • 通讯作者: 黎定军

Sub-cloning of Arabidopsis FLS2 and LysM Gene and Their Expression inTransgenic Tomatoes

CHEN Wu1;YANG Yu-ting1;LI Ying2;HUANG San-wen2;LIDing-jun1*   

  1. (1College of Bio-safety Science and Technology, Hunan Agricultural University, Changsha 410128, China; 2Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081,China)
  • Received:2009-04-27 Revised:2009-06-05 Online:2009-11-25 Published:2009-11-25
  • Contact: LIDing-jun1

摘要: 从拟南芥JAtYTAC基因组文库中完整地亚克隆了FLS2LysM 两个抗病基因, 亚克隆的DNA
片段中分别包含各自的启动子、内含子和终止子。利用农杆菌介导的方法分别将这两个基因转化番茄品种‘Moneymaker’, 转基因植株经PCR 初步筛选后再分别提取总RNA 进行RT-PCR, 以验证其是否在‘Moneymaker’中表达。试验结果表明, FLS2LysM 均能成功地在‘Moneymaker’中表达, 说明番茄的转录系统不仅能识别这两个抗病基因的启动子并起始转录, 还能正确识别并剪切这两个基因的外显子和内含子。据此推测, 拟南芥和番茄这两个远缘物种的转录机制存在相对保守的成份, 这为探索在这两个物种间R基因的功能及其下游信号通路的保守性提供了有价值的参考。

关键词: 拟南芥, 番茄, FLS2, LysM, 亚克隆, 表达

Abstract: FLS2 and LysM genes, two receptor-like kinases (RLKs) genes, containing their own promoters, introns and transcription terminators were sub-cloned from Arabidopsis TAC library, and then were transformed into‘Moneymaker’by Agrobacterium-mediated approach. Transgenic plants were identified by
polymerase chain reaction ( PCR). RT-PCR was performed to analyze two genesp expression in transgenic plants. The results illustrated that both FLS2 and LysM genes can be transcribed with the recognition of promoters, the splicing of introns and exons by tomato transcription system. It can be deduced that the transcription mechanisms of Arabidopsis and tomato could be conservative to some extent. This study provides a theoretical base for understanding functional conservation of R genes between Arabidopsis and tomato.

Key words: Arabidopsis thaliana, tomato, FLS2, LysM, sub-cloning, expression

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