膜荚黄芪香豆酸辅酶A连接酶基因(4CL)的克隆与表达分析

Cloning and expression of 4CL gene in Astragalus membranaceus

香豆酸辅酶A连接酶(4-coumarate:CoA lig-ase,4CL)是植物类黄酮类化合物和木质素等生物合成途径的一种关键酶,参与合成多种次生代谢物,在植物生长发育中起着重要作用。该试验采用同源克隆法和RACE技术从膜荚黄芪RNA中克隆得到4CL基因。利用生物信息学软件和荧光定量PCR技术,对该基因的功能和特征进行分析。结果表明:4CL基因(GenBank登录号KY086284)的cDNA序列长度为1937 bp,其ORF编码546个氨基酸,属于疏水性蛋白。实时荧光定量PCR分析发现,Am 4 CL基因在根、茎、叶中的表达趋势与毛蕊异黄酮葡萄糖苷的含量基本一致,推测Am 4 CL基因在膜荚黄芪异黄酮生物合成中发挥着重要的作用。

4coumarate:CoA lig-ase(4CL)is a key enzyme in biosynthetic pathways such as plant flavonoids and lignin.It participates in the synthesis of many secondary metabolites and plays an important role in plant growth and development.In this experiment,4 CL gene was cloned from Astragalus membranaceus by homologycloning method and RACE technique.Using bioinformatics software and real-time fluorescence quantitative PCR technology,the function and characteristics of this gene were analyzed.The results showed that the cDNA sequence of the 4 CL gene from A.membranaceus(Am 4 CL,GenBank accession number KY086284)was 1937 bp with an ORF encoding a hydrophobic protein comprising of 546 amino acids.The expression trend of in roots,stems and leaves was consistent with the contents of calycosin-7OβD glucoside speculating that Am 4 CL plays an important role in the biosynthesis of isofavone in A.membranaceus.