地黄C3H基因的克隆及生物信息学分析  

Cloning and bioinformatics analysis of C3H gene in Rehmannia glutinosa

作　　者：周延清[1] 邵露营 郭萌萌 朱佳琳 ZHOU Yanqing;SHAO Luying;GUO Mengmeng;ZHU Jialin(College of Life Sciences,Henan Normal University,Xinxiang 453000,Henan,China)

机构地区：[1]河南师范大学生命科学学院,河南新乡453000

出　　处：《广西植物》2020年第9期1281-1287,共7页Guihaia

基　　金：国家级项目培育基金(2016PL11);河南省自然科学基金(182300410018);校内结余项目(5201049170556)。

摘　　要：香豆酸-3-羟化酶属于植物中最大的蛋白酶细胞色素P450家族之一,在植物生命活动中发挥着重要作用。为了解地黄香豆酸-3-羟化酶基因RgC3H合成毛蕊花糖苷的功能,该研究基于地黄代谢组学分析获得KEGG途径中的C3H,采用多重比对在NCBI中获得同源基因的一个保守序列,并基于该保守序列和地黄SRA数据库,采用电子克隆和RT-PCR克隆技术获得地黄C3H基因全长CDS(RgC3H),对其进行生物信息学分析。结果表明:RgC3H基因全长为1530 bp,且编码一个含509个氨基酸、分子量为57.91 kD、无信号肽的蛋白质;基于氨基酸序列的结构分析显示,RgC3H有一个保守区域-P450结构域;系统进化分析结果显示,RgC3H与芝麻和猴面花的C3H基因具有很高的同源性。上述结果为进一步研究RgC3H基因在地黄毛蕊花糖苷生物合成途径中的作用奠定了基础。Coumarin-3-hydroxylase belongs to the cytochrome P450 family,one of the largest protease families,and plays an important role in plants.However,its role for verbascoside biosynthesis remains deficient.In order to understand the role of P-coumarate 3-hydroxylase for verbascoside biosynthesis in Rehmannia glutinosa(RgC3H),C3H was mined from a candidate KEGG pathway based on the metabolomics analysis of R.glutinosa,and a conserved sequence of homologous gene was obtained in NCBI by multiple alignment.According to its fragment and SRA database of R.glutinosa,we cloned its full length CDS(RgC3H)by electro cloning and RT-PCR,and performed its bioinformatics analyses.The results were as follows:RgC3H gene was 1530 bp in length and encoded a 509-amino acid protein with a molecular weight of 57.91 kD and without signal peptide;According to its amino acid sequence and structural analysis,RgC3H contained one conserved domain-cytochrome P450 domain,suggesting that it belonged to cytochrome P450 family;Phylogenetic tree analysis showed that RgC3H had high homology with the C3H genes of Sesamum indicum and Erythranthe guttata.This study laid a foundation for further research on the role of RgC3H gene in the pathway of verbascoside biosynthesis in Rehmannia glutinosa.

关 键 词：地黄 C3H 基因 基因克隆 生物信息学分析 

分 类 号：Q943[生物学—植物学]