文献类型：期刊文献

中文题名：当归4-香豆酸辅酶A连接酶基因克隆与组织表达分析

英文题名：Cloning and tissue expression of 4-coumarate coenzyme A ligase gene in Angelica sinensis

作者：温随超[1];王引权[1];雒军[1];夏琦[1];樊秦[1];荔淑楠[1];王振恒[1]

第一作者：温随超

机构：[1]甘肃中医药大学,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2015

卷号：40

期号：24

起止页码：4824

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：MEDLINE(收录号：27245029);CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；PubMed;

基金：国家自然科学基金项目(81260616;81060327)

语种：中文

中文关键词：当归;4-香豆酸辅酶A连接酶;基因克隆;基因表达

外文关键词：Angelica sinensis; 4-coumarate coenzyme A ligase; gene cloning; gene expression

摘要：4-香豆酸辅酶A连接酶(4-coumarate coenzyme A ligase,4CL)为高等植物苯丙烷代谢途径关键酶之一,在当归阿魏酸的生物合成中起调控作用。研究利用同源克隆结合c DNA末端快速扩增(RACE)技术克隆当归4CL编码基因全长c DNA序列,并用实时荧光定量PCR(qRT-PCR)分析4CL基因在当归幼苗生长过程中根、茎、叶组织中的表达特征。结果表明,获得的4CL c DNA序列(在Gen Bank已注册,登录号为KT880508)全长1 815 bp,含有1个1 632 bp的完整开放阅读框(opening reading frame,ORF),编码544个氨基酸的多肽链。4CL基因在当归幼苗不同生长阶段的根、茎、叶组织中均有表达,随着苗龄增大,茎、叶中的相对表达量显著增加(P<0.05),而根中表达量变化不大,表明4CL基因在当归幼苗中的表达具有明显的时空性。该研究为进一步研究4CL基因的结构与功能,解析当归阿魏酸的合成代谢机制和时空调控的分子基础奠定了工作基础。
4-coumarate coenzyme A ligase is a key enzyme of phenylpropanoid metabolic pathway in higher plant and may regulale the biosynthesis of ferulic acid in Angelica sinensis. In this study, the homology-based cloning and rapid amplification of cDNA ends （RACE） technique were used to clone a full length cDNA encoding 4-coumarate eoenzyme A ligase gene （4CL） , and then qRT-PCII was taken for analyzing 4CL gene expression levels in the root, stem and root tissue at different growth stages of seedlings of A..~inen,sis. The results showed that a full-length 4CL cDNA （1 815 bp） was obtained（ GenBank accession number： KT880508） whieb shares all open reading frame （ORF） of 1 632 bp, encodes 544 amino acid polypeptides. We found 4CL gene was expressed in all tissues inclu- ding leaf, stem and root of seedlings of A. sinerrsis. The expressions in the leave and stem were increased significantly with the growth of seedlings of A. sinensis（P 〈 0. 05） , while it in the root showed little change. It indicates a time-space pattern of 4CL gene expres- sion in seedlings of A. sinensis. These findings will be useful for establishing an experiment basis for studying the structure and function of 4CL gene and elucidating mechanism of ferulic acid biosynthesis and space-time regulation in A. sinensis.