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园艺学报 ›› 2004, Vol. 31 ›› Issue (6): 811-813.

• 研究报告 • 上一篇    下一篇

黄瓜花叶病毒反义2b 基因构建及其转基因初步研究

庄 木;王晓武*;谢丙炎*;杨宝军;方智远;黄和艳;郭淑华

  

  1. (中国农业科学院蔬菜花卉研究所, 北京100081)
  • 收稿日期:2004-02-13 修回日期:2004-06-14 出版日期:2004-12-25 发布日期:2004-12-25

Construction and Transformation of An Antisense CMV 2b Gene

Zhuang Mu;Wang Xiaowu*;Xie Bingyan*;Yang Baojun;Fang Zhiyuan;Huang Heyan; Guo Shuhua
  

  1. (Laboratory of Biotechnology , Institute of Vegetables and Flowers , Chinese Academy of Agricultural Sciences , Beijing 100081 , China)
  • Received:2004-02-13 Revised:2004-06-14 Online:2004-12-25 Published:2004-12-25

摘要: 研究构建了CMV 反义2b 基因的植物表达载体pZM 612 , 通过农杆菌侵染的叶盘转化法导入烟草, 分子检测证实获得了转反义2b 基因的SR1 烟草转基因植株, 初步的CMV 攻毒试验表明反义2b 基因可介导对CMV 的抗性。在此基础上, 将pZM 612 转化番茄品种‘红玛213’, 获得了卡那霉素抗性苗, PCR检测表明得到了PCR 阳性转化株。

关键词: 黄瓜花叶病毒, 反义2b 基因, 转基因番茄

Abstract: pBI1212based vector pZM 612 containing an antisense full2length 2b gene of CMV2BG was constructed and introduced into Nicotiana tabaccum var. SR1 , via Agrobacterium tumef aciens2mediated leaf2disc transformation. The regenerated plants were screened by PCR , and further verified by PCR-Southern and one-step RT-PCR. The results indicated that antisense 2b gene of CMV2BG had been integrated into the tobacco chromosomes ,and expressed at transcriptional level . The resistance of transgenic plants was observed after viral infection , indicating the mediation of resistant to CMV2BGby the antisense 2b gene. The antisense 2b gene of CMV2BG (pZM 612) was also introduced into Lycopersicon escuentum var. Hongmanao 213 , and PCR positive transgenic tomato plants were obtained.

Key words: CMV, Antisense 2b gene, Transgenic tomato

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