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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (18): 174-178.doi: 10.11924/j.issn.1000-6850.2012-0488

所属专题： 生物技术； 油料作物； 园艺

• 生物技术科学 • 上一篇    下一篇

甘蓝型油菜iMyAP9、iMyAP12基因过表达载体的构建及转化拟南芥

张伟 尹峰 何雯 魏然 刘春林 阮颖   

  • 收稿日期:2012-02-20 修回日期:2012-03-21 出版日期:2012-06-25 发布日期:2012-06-25
  • 基金资助:

    国家自然基金面上项目

The Over-expression Vector Construction of BnaiMyAP9 and BnaiMyAP12 and Transformation of Arabidopsis thaliana

  • Received:2012-02-20 Revised:2012-03-21 Online:2012-06-25 Published:2012-06-25

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摘要：

诱导型黑芥子酶协助蛋白(iMyAP)往往与黑芥子酶和黑芥子酶结合蛋白一起以复合体的形式存在，对植物的防御系统有着重要的作用。为了研究黑芥子酶协助蛋白在植物响应非生物胁迫方面的功能，通过以pBI121为载体，分别构建了CaMV35S::iMyAP9和CaMV35S::iMyAP12，浸花法转化拟南芥，用卡那霉素对转化植株进行筛选，并对抗性苗进行PCR检测，初步得到67株转基因植株；提取转基因拟南芥中RNA进行RT-PCR表达分析，结果表明，iMyAP9基因和iMyAP12基因在随机挑选的转基因拟南芥中获得了有效的表达。拟南芥转基因植株的获得为进一步探讨iMyAP9基因、iMyAP12基因的生物学功能奠定了基础。

关键词: 整理潜力, 整理潜力

Abstract:

Inducible Myrosinase-associated Protein (iMyAP) usually occur in complexes with Myrosinases and Myrosinases-binding protein plays important roles in the plant’s defense system. To study the functions of iMyAP in abiotic stress responses. Here, we constructed two plant over-expression vectors CaMV35S::iMyAP9 and CaMV35S::iMyAP12. Genetic transformation of Arabidopsis was performed by floral dip method. The T0 seeds were screened on medium with kanamycin and the anti-kanamycin plants were tested by PCR to confirm if anti-Kan plants were transgenic plants. 67 transgenic plants were obtained. The results of RT-PCR showed that iMyAP9 and iMyAP12 genes efficiently expressed in the transgenic arabidopsis plants, which the results lay a foundation for further studying the biological function of iMyAP9 and iMyAP12 genes.