膜荚黄芪肉桂酸-4-羟化酶(C4H)基因克隆与序列分析

Cloning and sequence analysis of cinnamate acid-4-hydroxylase(C4H) gene from Astragalus membranaceus

肉桂酸-4-羟基化酶是苯丙烷途径中继L-苯丙氨酸解氨酶(PAL)之后的第2个关键酶。为了从分子水平上了解C4H基因的功能特性,本文从植物进化的角度出发,根据Genebank中已有的豆科植物C4H氨基酸保守序列为基础,设计引物,通过RT-PCR和cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)方法克隆膜荚黄芪C4H基因,对其序列进行分析,为进一步研究其功能奠定基础。

Cinnamic acid-4-hydroxylase（C4H,EC1.14. 13.11）is the second key enzyme in phenylpropanoid metabolic pathway after L-phenylalnine ammonialyase（PAL）. In order to understand the functional prop- erties of C4H gene, designed PCR primers that target evolutionarily conserved sequeces of C4H gene in plants and cloned full-length sequences of C4H gene in A. membranaceus by the technique of RT-PCR and RACE, analysised characteristic of sequence.