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目的克隆鱼腥草1-脱氧-D-木酮糖-5-磷酸还原异构酶(DXR)基因,并分析其差异表达。方法采用RT-PCR方法获得DXR基因c DNA序列,并对DXR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DXR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DXR基因长为1 416 bp,编码471个氨基酸。生物信息学预测DXR蛋白不含跨膜区,不含信号肽。DXR基因在鱼腥草的叶片中表达丰度最高,其次是地下茎,再次是地上茎,花中表达量最低。结论首次从鱼腥草中克隆了DXR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 the 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) gene of Houttuynia cordata Thunb and analyze its differential expression. Methods The cDNA sequence of DXR gene was obtained by RT-PCR. The physical and chemical properties, protein secondary structure and three-dimensional structure of DXR protein were predicted and the function of DXR protein was predicted. The real-time PCR method was used to detect the DNA sequence of DXR gene in fish Houttuynia underground stems, aboveground stems, leaves, flowers in the expression. Results The cloned DXR gene was 1 416 bp in length and encoded 471 amino acids. Bioinformatics predicts that the DXR protein does not contain the transmembrane region and contains no signal peptide. DXR gene in Houttuynia leaves the highest abundance expression, followed by underground stems, and then aboveground stems, flower expression in the lowest. Conclusion The DXR gene was cloned from Houttuynia for the first time, which laid the foundation for further elucidating the important role of this gene in the metabolic pathway of terpineoids in Houttuynia regia.