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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (10): 168-172.

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

白菜转脂蛋白-CaMBP10基因RNAi沉默体系的初步建立

张蕊 胡文全 谢万钦 李翠凤   

  • 收稿日期:2010-08-13 修回日期:2010-11-30 出版日期:2011-05-15 发布日期:2011-05-15
  • 基金资助:

    白菜转脂蛋白BP10抗病原真菌功能与机制的研究;植物转脂蛋白抗病原真菌机制的研究

Prelimilary establishment of RNAi -mediated lipid transfer protein- CaMBP10 gene silencing system in Chinese cabbage

  • Received:2010-08-13 Revised:2010-11-30 Online:2011-05-15 Published:2011-05-15

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摘要：

为了研究白菜转脂蛋白-CaMBP10的体内功能，构建含有该基因片段的RNAi质粒，以农杆菌介导法对大白菜‘津育60’进行遗传转化，通过对转基因植物在生物与非生物胁迫中表型变化的研究，确定其体内功能。本文报告RNAi质粒的构建、鉴定及其转化植株基因组DNA中外源片段的鉴定。DNA分析结果显示外源基因已成功插入了白菜基因组，RT-PCR检测结果显示转化植株内源CaMBP10基因的表达明显降低，为后续的转脂蛋白体内功能研究奠定了基础。

关键词: 水产养殖, 水产养殖, 外来物种, 长江下游流域, 调查研究

Abstract:

In this paper，a RNAi approach was used in down-regulated CaMBP10 gene expression to study its functions in vivo. A 273 bp fragment of CaMBP10 cDNA was cloned from Chinese cabbage “JINYU 60” and inserted into the RNAi vector PTCK303. According to the results of detections, sense and anti-sense of CaMBP10 fragments had been cloned into RNAi victor between the left and right of an intro from rice. The Chinese cabbage “JINYU 60” was transformed by Agrobacterium tumefaciens LBA4404 which harbored this anti-sense CaMBP10 gene. The result of RT-PCR analysis shows that the CaMBP10 gene expression was reduced in transgenic plants. This work would serve as a good base for following research.