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.
ABSTRACT
OBJECTIVE: To investigate the chemical constituents from the stems of Viola japonica var. stenopetala Franch. ex H. METHODS: The constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography, and preparative TLC. The structures were identified on the basis of spectral data and physiochemical characteristics. RESULTS: Fifteen compounds were isolated from 70% ethanol extract of Viola japonica var. stenopetala Franch. ex H. and identified as β-sitosterol (1), daucosterol (2), chlorogenic acid (3), 7-hydroxycoumarin (4), stigmastero-3-O-β-D-glucopyranoside (5), dehydrololiolide (6), kaempferol-7-O-β-D-glucopyranoside (7), characterizedas(+)-pinoresinol-O-β-D-glucopyranoside (8), 5, 7-dihydroxy-3, 6-dimethoxyflavone (9), apigenin-7-O-β-D-glucoside (10), chryseriol (11), β-amyrin (12), robinin (13), kaempferol-3, 7-di-O-α-L-rahmnoside(14), and solagenin-6-O-β-D-quinovopyranoside(15). CONCLUSION: Compounds 8 and 15 are isolated from the plants in Gnaphalium L. for the first time. Compounds 5, 6, 8, 11, 14, and 15 are isolated from this plant material for the first time.
ABSTRACT
Objective: Acetyl-CoA C-acetyltransferase (AACT) is the initial enzyme in the terpenoid biosynthesis pathway of mevalonate (MVA), two units of acetyl-CoA were catalyzed to acetoacetyl-CoA. To clone the full length cDNA of AACT gene and carry out the bioinformatics analysis and expression analysis in order to provide the basis on resolving the mechanism of biosynthesis for terpenoid secondary metabolites from Houttuynia cordata. Methods: The cDNA sequence of AACT gene was obtained from H. cordata by using RT-PCR strategy. And the different expression of AACT gene in the rhizomes, stems, leaves, and flowers of H. cordata was analyzed by fluorescent quantitative PCR. Results: The cDNA contains a 1 218 bp open reading frame and encodes a predicted protein of 405 amino acids. No transmembrane region and signal peptide were present in AACT protein by bioinformatics prediction. Relative real-time PCR analysis indicated that AACT gene showed the highest transcript abundance in the stems and rhizomes of H. cordata lower levels in the flowers and leaves, the values of them were 1.49, 0.96, 0.20, and 0.10, respectively. Conclusion: This AACT gene is cloned from H. cordata for the first time. The results will provide a foundation for exploring the mechanism of the gene in terpenoid biosynthesis and metabolism in H. cordata.
ABSTRACT
Objective: To clone the 3-hydroxy-3-methylglutaryl coenzyme A redutase (HMGR) gene from Sambucus chinensis and analyze the differential expression. Methods: The sequence of HMGR was cloned from S. chinensis by using RT-PCR strategy. The physical and chemical properties, secondary structure, and tertiary structure of the HMGR protein were forecasted and analyzed, and its structure and function were predicted. And the different expression of HMGR gene in the rhizome, stems, leaves, and flowers was analyzed by fluorescent quantitative PCR. Results: The cDNA contains a 1782 bp open reading frame and encodes a predicted protein of 593 amino acids. Two transmembrane regions and no signal peptide were present in HMGR. Relative real-time PCR analysis indicated that HMGR showed the higher transcript abundance in the flowers and aerial stems, and the lower levels in the rhizomes and leaves. Conclusion: This study clones and expression analyzes HMGR gene from S. chinensis for the first time. The results will provide a foundation for exploring the mechanism of terpenoid biosynthesis in S. chinensis.