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目的克隆接骨草Sambucus chinensis 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RT-PCR方法获得HMGR基因c DNA序列,并对HMGR蛋白进行理化性质、蛋白二级结构及三级结构预测分析,并预测该蛋白功能;利用实时荧光定量PCR方法检测HMGR基因在接骨草的根状茎、地上茎、叶、花中的表达情况。结果克隆获得的HMGR基因c DNA全长为1 626 bp,编码593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基因主要在接骨草的花和地上茎中表达较高,其他器官表达相对较低。结论首次从接骨草中克隆了HMGR基因,为进一步阐明该基因在接骨草萜类化合物代谢途径中的重要作用奠定基础。
Objective To clone the gene of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) from Sambucus chinensis and analyze its differential expression. Methods The cDNA sequence of HMGR gene was obtained by RT-PCR. The physico-chemical properties, protein secondary structure and tertiary structure of HMGR protein were predicted and the function of HMGR protein was predicted. The real-time PCR method was used to detect the HMGR gene expression in Jiebucao Rhizomes, aboveground stems, leaves, flowers in the expression. Results The full length cDNA of HMGR gene cloned was 1 626 bp, encoding 593 amino acids. Bioinformatics prediction HMGR protein contains two transmembrane regions, does not contain signal peptide. The HMGR gene is mainly expressed in the flowers and aboveground stems of Crassulaceae, while the expression of other organs is relatively low. Conclusion The HMGR gene was cloned for the first time from the genus Crataegus, which laid the foundation for further elucidation of the important role of this gene in the metabolism of terbuthyla terpenoids.