cDNA cloning, difference expression in various organs, and bioinformatic analysis of HMGS gene in Sambucus chinensis / 中草药

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.