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农学学报 ›› 2015, Vol. 5 ›› Issue (11): 91-96.doi: 10.11923/j.issn.2095-4050.cjas15010042

所属专题: 生物技术

• 农业工程 农业机械 生物技术 食品科学 • 上一篇    下一篇

玫瑰查尔酮合酶CHS 基因的克隆及生物信息学分析

齐 宇,赵兰勇   

  1. 山东农业大学林学院,山东农业大学林学院
  • 收稿日期:2015-01-31 修回日期:2015-03-10 接受日期:2015-03-24 出版日期:2015-11-19 发布日期:2015-11-19
  • 通讯作者: 赵兰勇 E-mail:489348594@qq.com
  • 基金资助:
    山东省农业良种工程项目“高档园林花卉新品种选育”(鲁科农字[2014]96 号)。

Cloning and Bioinformatics Analysis of Chalcone Synthase Gene in Rosa rugosa

  • Received:2015-01-31 Revised:2015-03-10 Accepted:2015-03-24 Online:2015-11-19 Published:2015-11-19

摘要: 本研究旨在克隆玫瑰查尔酮合酶CHS 基因并对其进行生物信息学分析,以期为利用基因工程技术改变玫瑰花色、培育玫瑰新品种奠定良好基础。以玫瑰品种‘唐红’(Rosa rugosa‘Tanghong’)为试材,采用RT-PCR 和RACE 方法从花瓣中获得了玫瑰查尔酮合酶(chalcone synthase,CHS)基因的cDNA 全长。经生物信息学分析,该基因全长1416 bp,开放阅读框1167 bp,编码389 个氨基酸,其蛋白的分子式为C1899H3043N505O562S18,相对分子质量为42.518 kDa,等电点为6.12。该蛋白不稳定系数低于40,为稳定性蛋白,无信号肽和剪切位点存在。其蛋白质二级结构主要由42.16%的α 螺旋、29.05%的随机卷曲、10.54%的β转角和18.25%的延伸链组成。系统进化分析表明,玫瑰CHS 基因与同科植物月季、草莓亲缘关系最近,与其他科及同科其他属植物的亲缘关系较远。本研究首次克隆了玫瑰CHS 基因,获得了其生物信息学相关信息,探明了CHS 基因在类黄酮生物合成过程中的重要作用,为改良玫瑰花色提供了理论依据。

关键词: 绿豆、播种期、产量, 绿豆、播种期、产量

Abstract: The purpose of this study was to clone the chalcone synthase gene from Rosa rugosa and conduct bioinformatics analysis of the gene related information, and to lay a good foundation for changing Rosa rugosa’s color and breeding new varieties by genetic engineering. In this work, a full- length cDNA sequence of chalcone synthase (CHS) gene was obtained from petals of Rosa rugosa‘Tanghong’using RT-PCR and RACE. With the bioinformatics analysis, the cloned CHS gene was 1416 bp in full length, containing 1167 bp opening reading frame (ORF), which encoded 389 amino acids, the predicted CHS protein’s molecular formula was C1899H3043N505O562S18, whose relative molecular mass was 42.518 kDa and isoelectric point was 6.12. The CHS protein’s instability index was computed to be 37.45, which classified the protein as stable. And The CHS protein had no signal peptide and splice site. Characteristic prediction results indicated that the secondary structure of CHS protein mainly included 42.16%α -helix, 29.05% random coil, 10.54%β -sheet and 18.25% extended chain. Phylogenetic analysis indicated that the cloned CHS gene shared the closest homology with the CHS from Rosa chinensis and Fragaria×ananassa, and had farther homology with other plants. The CHS gene was cloned for the first time, this study obtained its bioinformatics information, proved the importance of CHS gene in flavonoids biosynthesis, provided a theoretical basis for improving Rosa rugosa’s color.