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目的克隆陆英Sambucus chinensis 3-羟基-3-甲基戊二酰辅酶A合成酶(HMGS)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HMGS基因c DNA序列并对HMGS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RT-PCR方法检测了HMGS基因在陆英的根状茎、地上茎、叶、花中的表达情况。结果克隆获得的HMGS基因c DNA全长为1 401 bp,编码466个氨基酸。生物信息学预测HMGS蛋白无跨膜区,不含信号肽。HMGS基因主要在陆英的花和根状茎中表达较高,在叶中表达相对较低。结论首次从陆英中克隆了HMGS基因,为进一步阐明该基因在陆英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 synthetase (HMGS) from Sambucus chinensis and analyze its differential expression. Methods Real-time fluorescence quantitative PCR (RT-PCR) was used to obtain the HMGS gene cDNA sequence. The physical and chemical properties, protein secondary structure and three-dimensional structure prediction of HMGS protein were predicted and the function of HMGS protein was predicted. RT- HMGS gene in Luying rhizome, aboveground stems, leaves, flowers in the expression. Results The full length cDNA of HMGS gene cloned was 1 401 bp, encoding 466 amino acids. Bioinformatics predicts that the HMGS protein has no transmembrane domain and no signal peptide. The HMGS gene is mainly expressed in the flowers and rhizomes of Lu Ying and is relatively low in leaves. Conclusion The HMGS gene was cloned for the first time from Lu Yingzhong, which laid the foundation for further elucidation of the important role of this gene in the metabolism of terbutirin.