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药用植物三七通过甲羟戊酸途径(mevalonic acid pathway)合成三萜皂苷生物合成的前体。甲羟戊酸激酶(mevalonate kinase,MVK)则是甲羟戊酸合成途径中ATP依赖的限速酶之一。本研究根据课题组已获得的三七[Panax notoginseng(Burk.)F.H.Chen]转录组数据中的MVK1转录本序列,利用RT-PCR方法获得三七MVK1(PnMVK1)基因的全长cDNA序列;对PnMVK1蛋白进行理化性质、蛋白二级结构及三维结构预测分析,并预测了该蛋白的结构与功能;利用实时荧光定量PCR方法检测了PnMVK1基因在三七的根、茎、叶、花中的表达情况。序列分析表明,克隆获得的三七PnMVK1基因长为1 164 bp,编码387个氨基酸,GenBank登录号JQ957844。生物信息学预测PnMVK1蛋白不含跨膜区,不含信号肽,具有GHMP激酶等保守结构域。PnMVK1基因在三七的根中表达丰度最高。本研究成功克隆并分析了三七MVK1的全长序列,为进一步阐明三七三萜代谢途径奠定基础。
The medicinal plant Panax notoginseng synthesizes triterpene saponin biosynthesis precursors through the mevalonic acid pathway. Mevalonate kinase (MVK) is one of the ATP-dependent rate-limiting enzymes in the mevalonate synthesis pathway. In this study, the full-length cDNA sequence of PnMVK1 gene was obtained by RT-PCR based on the MVK1 transcripts of Panax notoginseng (Burk.) FHChen transcriptome obtained from the research group. PnMVK1 protein was predicted by physical and chemical properties, protein secondary structure and three-dimensional structure prediction, and the structure and function of PnMVK1 protein were predicted. The expression of PnMVK1 gene in roots, stems, leaves and flowers of Panax notoginseng was detected by real-time fluorescence quantitative PCR Happening. Sequence analysis showed that the PnMVK1 gene of Panax notoginseng was 1 164 bp in length and encoded a protein of 387 amino acids with GenBank accession number JQ957844. Bioinformatics prediction PnMVK1 protein does not contain the transmembrane region, does not contain the signal peptide, with conserved domains such as GHMP kinase. The PnMVK1 gene was most abundantly expressed in roots of Panax notoginseng. In this study, we cloned and analyzed the full-length sequence of Panax Notoginseng MVK1, which laid the foundation for further elucidating the metabolic pathway of notoginseng triterpene.