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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (12): 249-254.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

拟南芥植物型PEPC基因人工小RNA表达载体的构建与遗传转化

冯都华 王伏林 陈锦清   

  • 收稿日期:2011-01-10 修回日期:2011-03-07 出版日期:2011-05-25 发布日期:2011-05-25
  • 基金资助:

    国家自然科学基金

Construction of Plant-type Phosphoenolpyruate Carboxylase (PEPC) Gene Artificial miRNA Expression Vector and Transformation in Arabidopsis thaliana

  • Received:2011-01-10 Revised:2011-03-07 Online:2011-05-25 Published:2011-05-25

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摘要：

磷酸烯醇式丙酮酸羧化酶（PEPC）是植物中具有多种生理功能的酶。为探索拟南芥中植物型PEPC基因对模式植物拟南芥脂肪酸含量以及抗逆性等方面的影响，本文构建了同时敲除拟南芥Atppc1、Atppc2和Atppc3基因的人工小RNA（amiRNA）植物表达载体pFGC-amiAtppc123，经根癌农杆菌EHA105介导，用花序浸染法转化拟南芥，成功获得转基因植株。RT-PCR半定量分析表明人工小RNA在转化植株中成功进行了超量表达。该试验为分析拟南芥脂肪酸含量以及抗逆性方面提供了基础材料。

关键词: 初步鉴定, 初步鉴定

Abstract:

To investigate the functions of PEPCase in lipid content, fatty acid composition and the response to environmental stress in C3 plants, Arabidopsis thaliana plant-type PEPCase genes (referred to as Atppc1, Atppc2 and Atppc3) were silence by the artificial miRNA (amiRNA) technique. We constructed the plant expression vector named pFGC-amiAtppc123 and transformed it into Arabidopsis thaliana via in floral dip method. Transgenic plants were confirmed by PCR. The amiRNA transcript levels were detected by semi-quantitative RT-PCR. The results showed that amiRNA expression was drastically increased in transgenic plant compared with non-transgenic plants.