白木香肉桂酸-4-羟基化酶(C4H)基因的克隆及表达分析  

Cloning and expression analysis of cinnamate 4-hydroxylase(C4H)reductase gene from Aquilaria sinensis

作　　者：梁良[1] 韩晓敏[2] 张争[3,4] 郭庆梅[1] 徐艳红[3] 刘娟[3] 廖永翠[3] 

机构地区：[1]山东中医药大学药学院,山东济南250012 [2]燕山大学环境与化学工程学院,河北秦皇岛066004 [3]中国医学科学院北京协和医学院药用植物研究所,北京100193 [4]中国医学科学院北京协和医学院药用植物研究所海南分所,海南万宁571533

出　　处：《中国中药杂志》2014年第10期1767-1771,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(31000136;81173481);北京市自然科学基金项目(6102024);教育部新世纪优秀人才支持计划项目(2008);高等学校博士学科点专项科研基金项目(20091106120009);国家"十二五"科技支撑计划项目(2011BAI01B07);海南省中药现代化专项项目(2010ZY001;2012ZY002);海南省重大科技研发专项(ZDZX20100006)

摘　　要：对国产沉香的基原植物白木香Aquilaria sinensis肉桂酸-4-羟基化酶基因C4H编码区进行克隆,并对其进行生物信息学分析和表达分析。根据已报道的白木香伤害转录组高通量测序结果分析获得1条具有肉桂酸羟化酶酶保守结构域的C4H基因序列,采用RT-PCR技术克隆得到白木香C4H基因编码区序列,并对C4H蛋白进行生物信息学分析。另外,采用qRT-PCR方法对C4H基因在伤害胁迫下的表达模式进行分析。序列分析表明,所克隆的C4H基因编码区开放阅读框(opening reading frame,ORF)长为1 515 bp,编码514个氨基酸,GenBank登录号为KF134783。qRT-PCR实验证明C4H基因受不同伤害处理的诱导,分别在8,20 h达到最高表达水平。白木香C4H基因的编码区序列的分离克隆为进一步研究C4H蛋白在白木香中黄酮及芳香族类化合物合成途径中的功能及表达调控奠定基础。The study aimed to clone the open reading frame of cinnamate 4-hydroxylase（C4H） from Aquilaria sinensis and analyze the bioinformatics and expression of the gene.One unique sequence containing C4H domain was discovered in our previous reported wound transcriptome dataset of A.sinensis.The open reading frame of C4H was cloned by RT-PCR strategy with the template of mixed RNA extracted from A.sinensis stem which treated by different wound time.The bioinformatic analysis of this gene and its corresponding protein was performed.C4H expression profiles in responds to MeJA（ methyl jasmonate） application were analyzed by realtime PCR.The length of C4H open reading frame（ORF） was 1 515 bp,encoding 514 amino acids.The GenBank accession number is KF134783.Inducible-experiments showed that the genes were induced by mechanical wound as well as MeJA induction,and reached the highest expression level at 8 h and 20 h,respectively.The full-length cDNA of C4H and its expression patterns will provide a foundation for further research on its function in the molecular mechanisms of aromatic compounds and flavonoids biosynthesis.

关 键 词：C4H酶 白木香 芳香族化合物 伤害 

分 类 号：S567.19[农业科学—中草药栽培]