烟草1-脱氧木酮糖-5-磷酸合成酶(DXS)基因全长cDNA的克隆及表达分析
Molecular Cloning and Expression Analysis of the Full-Length cDNA of 1-Deoxy-D-Xylulose-5-Phosphate Synthase (DXS) Gene from Cultivated Tobacco (Nicotiana tabacum L.)
摘要: 利用反转录聚合酶链式反应(RT-PCR)结合快速扩增cDNA末端(RT-PCR)技术克隆了烟草1-脱氧木酮糖-5-磷酸合成酶(DXS)基因的全长cDNA(GenBank accession number: CBA12009)。序列分析表明该基因全长2386 bp，含有一个编码718个氨基酸残基的开放阅读框(ORF)。根据cDNA序列预测的蛋白质的理论等电点和分子量分别为7.31和77.413 kDa。该蛋白与辣椒和番茄的DXS的氨基酸序列的同源性分别达到96%和95%。对来自10种植物(包含烟草)的DXS的聚类分析显示，烟草的DXS与辣椒和番茄的DXS同属一个支系。Northern杂交分析显示，DXS基因在叶片和幼茎中表达水平最高，而在根和花器官中表达水平很低，这表明该基因的产物可能在绿色组织中参与次生物质的代谢。该基因cDNA序列的获得为通过基因工程途径改良烟叶香气品质奠定了基础。
Abstract: 1-deoxy-D-xylulose-5-phosphate synthase (DXS) gene from Nicotiana tabacum was cloned by using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Rapid Amplification of cDNA Ends (RACE) strategies. The complete cDNA sequence of DXS is 2386 bp in length, and contains an open reading frame (ORF) encoding 718 amino acid residues (GenBank accession number: CBA12009). The pI/MW (molecular weight) of the deduced protein is 7.31 and 77.413 kDa. The protein is most similar to DXS of Capsicum annuum with the highest amino acid identity of 96%, and to DXS of Solanum lycopersicum with the next highest amino acid identity of 95%. Phylogenetic analysis constructed with 10 plant DXSs showed that the deduced DXS formed a cluster with those from C. annuum and S. lycopersicum. Northern blot analysis revealed N. tabacum DXS gene is predominantly expressed in leaf and stem, and expressed at a very low level in root and flower, suggesting that the gene product is involved in metabolism of allelochemicals in green tissues. This work laid a foundation for improving the quality of tobacco fragrance by transgene in the near future. 
文章引用：章娟娟, 陈小艺, 方君, 杨之帆. 烟草1-脱氧木酮糖-5-磷酸合成酶(DXS)基因全长cDNA的克隆及表达分析[J]. 植物学研究, 2018, 7(4): 367-374. https://doi.org/10.12677/BR.2018.74045

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