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葡萄中两个编码 3-脱氧-D-阿拉伯庚酮糖-7-磷酸合酶同工基因的分子特征和表达分析。

Molecular characterization and expression analysis on two isogenes encoding 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase in grapes.

机构信息

Centre for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.

出版信息

Mol Biol Rep. 2011 Oct;38(7):4739-47. doi: 10.1007/s11033-010-0611-3. Epub 2010 Dec 4.

Abstract

3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHPS) is an entry enzyme of the shikimate pathway that connects primary carbohydrate metabolism with the biosynthesis of most secondary metabolites in plants. In the present study, two DAHPS cDNAs were cloned from grape berries (Vitis vinifera) and designated as VvDAHPS-1 and VvDAHPS-2. These two cDNA sequences share 75.7% of the identities. Their DNA corresponding to the two isogenes both contain four introns. The deduced proteins from two cDNAs had different NH(4)-terminal regions and putative mature regions sharing sequence, molecular size and pI value similarity. Both of VvDAHPSs had a close evolution relationship with Populus trichocarpa DAHPSs. The prokaryotically-expressed VvDAHPSs both manifested DAHPS catalytic activity and Mn(2+)-activated effects. Analysis by real time-PCR showed that VvDAHPS-1 and VvDAHPS-2 were expressed in all the tested tissues, but their expression patterns accompanying with berry mature varied in the skin, pulp and seeds. The results give new insight into further study on regulatory mechanism of grape phenolics biosynthesis.

摘要

3-脱氧-D-阿拉伯庚酮糖-7-磷酸合酶(DAHPS)是连接初级碳水化合物代谢与植物中大多数次生代谢物生物合成的莽草酸途径的入口酶。在本研究中,从葡萄浆果(Vitis vinifera)中克隆了两个 DAHPS cDNA,并分别命名为 VvDAHPS-1 和 VvDAHPS-2。这两个 cDNA 序列具有 75.7%的同一性。它们的两个基因座的 DNA 均包含四个内含子。两个 cDNA 推导的蛋白质具有不同的 NH4-末端区域和假定的成熟区域,具有序列、分子大小和 pI 值相似性。两个 VvDAHPSs 与毛白杨 DAHPSs 具有密切的进化关系。原核表达的 VvDAHPSs 均表现出 DAHPS 催化活性和 Mn2+激活效应。实时 PCR 分析表明,VvDAHPS-1 和 VvDAHPS-2 在所有测试的组织中均有表达,但它们在果皮、果肉和种子中伴随浆果成熟的表达模式不同。这些结果为进一步研究葡萄酚类生物合成的调控机制提供了新的见解。

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