桂花查耳酮合酶chs基因的克隆及序列分析
Cloning and Sequencing Analysis of Osmanthus fragrans Chalcone Synthase Gene
徐秀荣 1马燕 1臧德奎1
作者信息
- 1. 山东农业大学林学院,山东泰安271000
- 折叠
摘要
克隆与桂花(Osmanthus fragrans)花色形成相关基因查耳酮合酶(Ofchs),并对该基因的序列特征、进化关系进行分析,为探究该基因在桂花花色形成中的功能提供理论依据.以桂花‘丹桂’品种为试材,采用RT-PCR和RACE方法克隆丹桂查耳酮合酶(chalcone synthase,chs)基因eDNA全长.利用NCBI和DNAMAN分析软件,对该基因进行核苷酸序列以及氨基酸序列的分析,利用DNAMAN软件直接构建系统进化树,以分析该基因在不同物种间的进化关系.结果表明:从‘丹桂’花瓣中成功获得了1个丹桂查耳酮合酶基因cDNA全长,该基因全长为1383 bp,包含48bp的5'非编码区、162 bp的3'非编码区和1个长度为1173 bp编码390个氨基酸的开放阅读框.序列分析表明,该基因具有Cys164、His303和Asn336 3个活性位点及2个决定底物特异性的苯丙氨酸残基Phe215、Phe265.该基因也具有查尔酮合酶家族的2条特征多肽序列:RFMMYQQGCFAGGTVLR和GVLFGFGPGL.Blast序列分析显示,该基因与同属木犀科的油橄榄相似性达95％,与其他双子叶植物的相似性达72％～76％.克隆出的丹桂chs基因属于查尔酮合酶家族,系统进化分析表明,该基因聚类具有明显的种属特性,桂花与唇形科等植物的chs亲缘关系较近,相似性达76％以上,符合植物分类学分类,为今后探究该基因在花色行形成中的作用机制奠定了基础.
Abstract
The full-length cDNA of chs gene of Osmanthus fragrans was cloned which was related to color formation, the characteristics of the gene sequence and evolution relationships were analyzed to provide a theoretical basis for exploring gene function in the form of color.In this study, a full-length cDNA sequence of chs gene was obtained from Osmanthusfragrans by using RT-PCR and RACE.The gene nucleotide and amino acid sequences were analyzed by using NCBI and DNAMAN and a phylogenetic tree was constructed by using DNAMAN software directly, which would help analyze the gene evolutionary relationships among different species.Result showed that: the full-length gene was cloned from Osmanthus fragrans successfully, which was displayed as Osmanthus fragrans chs gene, which was 1383 bp in full length and included 5'URT of 48 bp and 3' URT of 162 bp, had an open reading frame of 1173 bp encoding a deduced polypeptide of 390 amino acids.Sequence analysis revealed that this gene had 3 active sites: Cys164, His303 and Asn336, and 2 decisions substrate specificity of phenylalanine residues: Phe215 and Phe265 Ofchs had 2 characteristics of polypeptide sequence of chs synthase superfamily: RFMMYQQGCFAGGTVLR and GVLFGFGPGL.Blast sequence showed that similarity between Osmanthu fragrans and Olea europaea was up to 95％ and was at around 72％-76％ with the other dicots.Ofchs gene belonged to chs family, phylogenetic analysis showed that the gene cluster had the distinct species characteristics, Osmanthus fragrans had a near relationship to Labiatae, which was up to 76％,and it was in line with plant taxonomy, and laid the foundation to explore the mechanism of Ofchs in the formation of the color line for the future.
关键词
桂花/查耳酮合酶基因/克隆/序列分析Key words
Osmanthus fragrans/chs/cloning/sequencing analysis引用本文复制引用
基金项目
山东省自然科学基金(ZR2013CQ008)
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据