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目的克隆鱼腥草1-脱氧-D-木酮糖-5-磷酸合成酶1(DXS1)基因并分析其表达差异。方法根据已经获得的鱼腥草DXS1转录本序列设计1对引物,采用RT-PCR方法获得DXS1基因cDNA序列并对DXS1蛋白进行理化性质、蛋白二级结构及三维结构预测分析,并预测了该蛋白功能;利用实时荧光定量PCR方法检测了DXS1基因在鱼腥草的地下茎、地上茎、叶、花中的表达情况。结果克隆获得的DXS1基因长为2 172 bp,编码723个氨基酸。生物信息学预测DXS1蛋白不含跨膜区,不含信号肽,具有定位肽。DXS1基因在鱼腥草的花中表达丰度最高,其次是叶片,再次是地下茎,地上茎中表达量最低。结论首次从鱼腥草中克隆了DXS1基因,为进一步阐明该基因在鱼腥草萜类化合物代谢途径中的重要作用奠定基础。
Objective To clone and analyze the gene expression of 1-deoxy-D-xylulose-5-phosphate synthase 1 (DXS1) in Houttuynia cordata Thunb. Methods A pair of primers was designed according to the DXS1 transcript sequence of Houttuynia cordata. The cDNA sequence of DXS1 gene was obtained by RT-PCR and the physicochemical properties, secondary structure and three-dimensional structure of DXS1 protein were predicted and predicted. The expression of DXS1 in underground stem, aboveground stem, leaf and flower of Houttuynia cordata was detected by real-time fluorescence quantitative PCR. Results The cloned DXS1 gene was 2 172 bp in length and encoded 723 amino acids. Bioinformatics prediction DXS1 protein does not contain the transmembrane region, does not contain the signal peptide, with positioning peptide. The expression of DXS1 gene in the flowers of Houttuynia cordifolia was the highest, followed by the leaves, again the underground stems, and the lowest in the shoots. Conclusion The DXS1 gene was cloned from Houttuynia for the first time, which laid the foundation for further elucidation of the important role of this gene in the metabolic pathway of the genus Houttuynia.