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目的克隆鱼腥草3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)基因并分析其差异表达。方法采用实时荧光定量PCR(RT-PCR)方法获得HMGR基因cDNA序列并对HMGR蛋白进行理化性质、蛋白二级结构及三维结构预测分析,并预测该蛋白功能;利用RT-PCR方法检测HMGR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HMGR基因cDNA全长为1 626 bp,编码541个氨基酸。生物信息学预测HMGR蛋白含2个跨膜区,不含信号肽。HMGR基因主要在鱼腥草的花中表达,其他器官中表达相对较低,地下茎中表达量最低。结论首次从鱼腥草中克隆了HMGR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作用奠定基础。
Objective To clone the gene of Houttuynia cordata 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and analyze its differential expression. Methods The cDNA sequence of HMGR gene was obtained by real-time fluorescence quantitative PCR (RT-PCR) and the physicochemical properties, protein secondary structure and three-dimensional structure prediction of HMGR protein were predicted and the function of HMGR protein was predicted. RT- Houttuynia underground stems, aboveground stems, leaves, flowers in the expression. Results The full-length cDNA of HMGR gene cloned was 1 626 bp, encoding 541 amino acids. Bioinformatics prediction HMGR protein contains two transmembrane regions, does not contain signal peptide. The HMGR gene is mainly expressed in flowers of Houttuynia, the expression is relatively low in other organs, and the expression level in underground stems is the lowest. Conclusion The HMGR gene was cloned from Houttuynia for the first time, which laid the foundation for further clarifying the role of this gene in the metabolic pathway of the genus Houttuynia.