Feng Xu, Shui-Yuan Cheng, Shu-Han Cheng, Yan Wang, He-Wei Du
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引用次数: 0
摘要
查尔酮合成酶(Chalcone synthase, CHS)是黄酮类化合物生物合成途径的第一步和关键调控步骤。利用cDNA末端快速扩增法(RACE)从银杏叶中分离到一个查尔酮合成酶基因。全长cDNA,编号为GbCHS2,全长1608 bp (GenBank登录号:DQ054841),包含1173 bp的开放阅读框,编码391个氨基酸的蛋白。结果表明,预测的GbCHS2氨基酸序列与GbCHS1具有较高的序列相似性。GbCHS1蛋白的所有活性位点和活性位点基序也在GbCHS2中发现。银杏叶片生长过程中CHS活性与黄酮类物质积累的相关分析表明,CHS可能是银杏叶片黄酮类物质生物合成途径中的限速酶。半定量RT-PCR分析结果显示,银杏叶片中黄酮类物质积累与chs基因的转录水平变化平行,表明chs基因是调控银杏叶片中黄酮类物质积累的特定关键基因。
Time course of expression of chalcone synthase gene in Ginkgo biloba.
Chalcone synthase (CHS) catalyses the first and key regulatory step of flavonoid biosynthetic pathway. A chalcone synthase gene was isolated from Ginkgo biloba leaves using the method of rapid amplification of the cDNA ends (RACE). The full-length cDNA, designated as GbCHS2, is 1,608 bp in length (GenBank accession No. DQ054841) and contains an open reading frame of 1,173 bp encoding a protein of 391 amino acids. Alignment of the predicted amino acid sequence of GbCHS2 has been shown to have high sequence similarity with GbCHS1. All the active sites and active site motifs in GbCHS1 protein were also found in GbCHS2. Correlation analysis between CHS activity and flavonoid accumulation during ginkgo leaf growth indicated that CHS might be the rate-limiting enzyme in the biosynthesis pathway of flavonoids in ginkgo leaves. Results of semi-quantitative RT-PCR analysis showed that flavonoid accumulation paralleled the transcription level of change in chs gene, suggesting chs gene as the specific key gene regulating flavonoid accumulation in ginkgo leaves.