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| 拟南芥光敏色素A反义RNA载体的构建及转化 |
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王 海1,2,3, 王小兰2, 周玉萍2, 段 俊1, 田长恩2*
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1.中国科学院 华南植物园, 广州 510650;2.广州大学 植物抗逆基因功能研究广州市重点实验室,
广州 510006;3.中国科学院 研究生院, 北京 100049
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| 摘要: |
| 光敏色素A是植物远红光信号的关键受体,在植物光信号转导途径中起重要作用。一些具有重要功能的基因(如生长素反应因子8:auxin response factor 8,ARF8)受到PhyA调控。本实验拟通过构建PhyA基因反义株系,研究PhyA调控ARF8等重要功能基因表达的机理。以培养7 d的拟南芥幼苗为材料提取RNA,利用RT-PCR技术扩增出PhyA的全长CDS,将其反向插入表达载体pMD1得到反义表达载体pMD1-PhyA CDSR,并通过农杆菌介导转化拟南芥,经卡那霉素抗性平板筛选和PCR鉴定得到13个PhyA转基因株系。pMD1-PhyA CDSR及其转基因株系的获得为研究PhyA调控ARF8等基因表达的机理奠定了材料基础。 |
| 关键词: 光敏色素A 反义表达载体 拟南芥 生长素反应因子8 |
| DOI： |
| 分类号:Q789 |
| Fund project: |
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| Construction and transformation of antisense RNA expression vector of Arabidopsis phytochrome A gene |
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WANG Hai1,2,3, WANG Xiao-Lan2, ZHOU Yu-Ping2,
DUAN Jun1, TIAN Chang-En2*
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1.South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China;2.Guangzhou Key
Laboratory for Functional Study on Plant Stress-Resistant Genes, Guangzhou University, Guangzhou 510006,
China;3.Graduate University of Chinese Academy of Sciences, Beijing 100049, China
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| Abstract: |
| Phytochrome A(PhyA)is the specific photoreceptor which receives far-red light in plants. Many critical genes,including auxin response factor 8(ARF8),are regulated by PhyA. Therefore,it is necessary to construct the PhyA-antisence transgenic lines for revealing the mechanism underlying phyA-regulated genes expression. The PhyA CDS was amplified from Arabidopsis total RNA by RT-PCR method. Then,it was inserted reversely into pMD1 to get the antisence expression vector pMD1-PhyA CDSR. The transgenic lines were acquired by agrobacterium-mediated method. 13 transgenic lines have been acquired by Kmr screening and PCR analysis. The vector pMD1-PhyA CDSR and the transgenic lines will be useful to study how the genes,such as ARF8,are regulated by PhyA. |
| Key words: phytochrome A antisense expression vector Arabidopsis auxin response factor 8 |
