| 烟草红素氧还蛋白基因NtRD1的克隆及其RNAi载体构建 |
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| 引用本文： | 冯仁军,卢利方,程萍,袁克华,张银东.烟草红素氧还蛋白基因NtRD1的克隆及其RNAi载体构建[J].热带作物学报,2008,29(3):358-362. |
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| 作者姓名： | 冯仁军 卢利方 程萍 袁克华 张银东 |
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| 作者单位： | 中国热带农业科学院热带生物技术研究所,海口,571101;海南大学农学院,海口,570228 |
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| 基金项目： | 中央级公益性科研院所基本科研业务费项目资助 |
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| 摘 要： | 采用RACE(Rapid amplification of cDNA ends)技术进行烟草红素氧还蛋白基因NtRD1(Nicotianatabacum rubredoxin1)全长cDNA的克降,获得了全长808 bp的cDNA,该cDNA具有完整的开放阅读框架,编码204个氨基酸,其中包含一个保守的红素氧还蛋白结构域.同源分析结果表明,NtRD1与其它植物已克隆的RDs具有较高的一致性.系统发育分析结果表明,NtRD1与其它物种RDs的亲缘关系由近及远依次是马铃薯、水稻、葡萄、拟南芥和藻类.根据RNAi(RNA interference)载体构建原则,成功地构建了干扰NtRD1的RNAi植物表达载体pCAMBIA1301-NtRD1-RNAi,为进一步探明该基因在烟草中的功能奠定了基础.
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| 关 键 词： | 烟草 RACE 全长cDNA NtRD1 RNAi |
Cloning of Rubredoxin Gene NtRD1 in Nicotiana tabacum and Construction of Its Corresponding RNAi Vector |
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| Feng Renjun,Lu Lifang,Cheng Ping,Yuan Kehua and Zhang Yindong.Cloning of Rubredoxin Gene NtRD1 in Nicotiana tabacum and Construction of Its Corresponding RNAi Vector[J].Chinese Journal of Tropical Crops,2008,29(3):358-362. |
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| Authors: | Feng Renjun Lu Lifang Cheng Ping Yuan Kehua and Zhang Yindong |
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| Institution: | 1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences;2.College of Agronomy, Hainan University;1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences;2.College of Agronomy, Hainan University;1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences;2.College of Agronomy, Hainan University;1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences;2.College of Agronomy, Hainan University;1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences;2.College of Agronomy, Hainan University |
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| Abstract: | RACE strategy was used to clone full-length cDNA of rubredoxin(RD) gene in Nicotiana tabacum (Nt). An 808bp-length cDNA of NtRD1 with a completed open reading frame (ORF) was cloned, and the ORF encoded 204 amino acids containing a conserved rubredoxin domain. Homology analysis indicated that NtRD1 shared high identity with the reported RD of other plants. Phylogenetic analysis showed that the systematic relationships from the close to the far between the NtRD1 and the RDs of other species were Solanum tuberosum, Oryza sativa, Vitis vinifera, and Arabidopsis thaliana and algae in order. According to the principles of RNAi vector construction, plant expression vector pCAMBIA1301-NtRD1-RNAi was successfully constructed, and it could express dsRNA of NtRD1. All these have established a base for further research of the gene function in tobacco. |
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| Keywords: | tobacco RACE full-length cDNA NtRD1 RNAi |
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