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[摘要]
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cDNA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cDNA全长为1401 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。
[Key word]
[Abstract]
Objective To clone 3-hydroxy-3-methylglutaryl coenzyme A synthetase (HMGS) gene from Sambucus chinensis and analyze the difference expression. Methods The sequence of HMGS gene was cloned from S. chinensis by using RT-PCR strategy. The physiochemical properties, secondary structure, and three-dimensional structure of HMGS protein were forecasted and analyzed, and its structure and function were predicted. And the difference expression of HMGS gene in the rhizome, stems, leaves, and flowers of S. chinensis was analyzed by fluorescent quantitative PCR. Results The cDNA contains a 1 401 bp open reading frame and encodes a predicted protein of 466 amino acids. No transmembrane region and no signal peptide were present in HMGS protein. Relative real-time PCR analysis indicated that HMGS gene showed the higher transcript abundance was in the flowers and rhizomes, while was lower in the leaves. Conclusion The HMGS gene is first cloned from S. chinensis and the result will provide a foundation for elucidating the mechanism of the gene in the metablism pathway of terpenoid in S. chinensis.
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[基金项目]
湖南省科技计划项目(2011NK3045,2013FJ6090);湖南省生物类专业大学生创新训练中心资助