苎麻C3H基因的克隆、生物信息学分析及表达分析

Gene Cloning, Bioinformatics and Expression Analysis of C3H Gene from Boehmeria nivea L.

旨在为后期验证C3H基因编码蛋白功能并最终实现低木质素优良苎麻品种的基因工程育种奠定基础。基于苎麻高通量测序信息,利用RACE技术克隆C3H基因全长序列,对其进行生物信息学分析;并利用荧光定量技术对苎麻C3H的表达模式进行研究。获得了全长为1940 bp的苎麻C3H基因c DNA序列Bn C3H-1(Gen Bank登录号为KY078743)。Bn C3H-1基因编码蛋白理化性质和结构的分析表明,该基因编码一个含511个氨基酸,分子量为57.80 k D的亲水性蛋白,它可能定位在内质网上。氨基酸序列和结构分析显示Bn C3H-1有一个保守区域,即P450结构域。系统进化分析表明,该基因与巨桉和苦荞C3H基因具有很高的同源性。

The aim is to provide a basis for verifying the protein function of coumarate 3-hydroxylase gene （C3H） in the later period, and improving the genetic engineering breeding of Boehmeria nivea variety with low content lignin. Based on the information of high-throughput sequencing of Boehmeria nivea, the full-length sequence of C3H was cloned by the RACE technology, and then the bioinformatics analysis was conducted. The expression pattern of C3H was tested respectively by fluorescent quantitative technology. BnC3H-1 （cDNA sequence of C3H） with 1940 bp in length was obtained （GenBank accession number： KY078743）. The analysis of the physical and chemical properties and structure of BnC3H-1 gene encoding protein showed that the gene encoded a hydrophilic protein containing 511 amino acids with a molecular weight of 57.80 kD, it might be localized in the endoplasmic reticulum. According to the sequence of amino acid and structural analysis, BnC3H-1 contained one conserved domain, cytochrome P450 domain. Phylogenetic analysis showed that the gene had high homology with C3H genes of Eucalyptus grandis and Fagop3,rum tataricum.