紫苏肉桂酸4-羟基化酶基因的克隆与表达  

Cloning and expression analysis of cinnamate 4-hydroxylase gene from Perilla frutescens

作　　者：宋西红[1] 郝磊[1] 吕晓玲[1] 郭宇[1] 魏曼[1] 赵辰辰[1] 

机构地区：[1]天津科技大学食品工程与生物技术学院,天津300457

出　　处：《广东农业科学》2015年第11期124-129,共6页Guangdong Agricultural Sciences

基　　金：国家"863"计划项目(2007AA100401)

摘　　要：为阐述紫苏中迷迭香酸合成的分子调节机制,应用同源克隆和RACE技术首次从紫苏叶片中克隆得到肉桂酸-4-羟基化酶基因(Per C4H)的全长c DNA序列(Gen Bank登录号:KM434189)。Per C4H基因总长度为1 667 bp,基因的开放阅读框(ORF)为1 518 bp,编码505个氨基酸残基,还含有长度为21 bp的5′非编码区(5′UTR)和95 bp的3′非编码区(3′UTR)。系统进化树分析得到Per C4H基因与唇形科植物丹参和黄芩的亲缘性最近。由实时荧光定量PCR分析可知,该基因在紫苏中具有组织表达特异性,根中的表达最高,茎中次之,而叶中最少。一定浓度的赤霉素与茉莉酸甲酯能提高Per C4H的表达量,而脱落酸和黑暗处理能降低Per C4H的表达量。In order to clarify the molecular mechanism of romarinic acid （RA） biosynthesis, the cDNA of cinnamate 4-hydroxylase （PerC4H）gene was firstly cloned from the leaves of Perilla frutescens （GenBank accession No： KM434189） by homology-based cloning and rapid amplification of cDNA ends （RACE） technique. The full-length of PerC4H cDNA was 1 667 bp, containing 1 518 bp open reading frame （ORF）which encoded a pepitide of 505 amanio acids and two non-coding regions, 21 bp 5＇ UTR and 95 bp 3＇ UTR. Phylogenetic tree analysis showed that PerC4H had the closest relationship with Salvia miltiorrhiza and Scutellaria balcalensis. Real time quantitative PCR showed that expression of PerC4H was the hishest in root, moderate in stem, and the lowest in leaf. Further expression analysis showed that Gibberellins and MeJA treatments could up-regulate the transcript level of PerC4H, while ABA and dark treatments could down-regulate its transcription level in some degree.

关 键 词：紫苏 迷迭香酸 肉桂酸4-羟基化酶 基因克隆 表达分析 

分 类 号：Q786[生物学—分子生物学]