玫瑰查尔酮合酶CHS基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Chalcone Synthase Gene in Rosa rugosa

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c DNA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 k Da,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。

The purpose of this study was to clone the chalcone synthase gene from Rosa rugosa and conductbioinformatics analysis of the gene related information, and to lay a good foundation for changing Rosa rugosa’scolor and breeding new varieties by genetic engineering. In this work, a full- length c DNA sequence ofchalcone 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 openingreading frame（ORF）, which encoded 389 amino acids, the predicted CHS protein’s molecular formula wasC1899H3043N505O562S18, whose relative molecular mass was 42.518 k Da 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 secondarystructure 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 genewas 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.