山西农业大学学报（自然科学版）2017，Vol.37Issue(11)：767-773,7.
苦荞C4H基因的cDNA克隆及生物信息学分析
Cloning and bioinformatics analysis of C4H gene in Tatary Buckwheat
摘要
Abstract
[Objective]In order to understand the structural characteristics and phylogenetic evolution of the key enzyme—cinnamate-4-hydroxylase gene in plant phenylpropane metabolic pathway,we wanted to clone the cinnamate-4-hydroxylase (C4H)gene and analyze it by bioinformatics.[Methods]In this study,we used RT-PCR to clone the cDNA and DNA sequences of C4 H gene from tartary buckwheat,the genetic structure and protein physicochemical properties were analyzed by bioinformatics,and constructed the phylogenetic tree by maximum likelihood method . [Result]The cDNA sequences of the two genes were 1812 bp and 1476 bp,and respectively encoding 504 and 491 a-mino acid residues.The DNA sequence of two genes were 2774 bp and 2364 bp,including three exons and two in-trons,there were significant differences in the first exon and two introns sequence length.Protein molecular weight were 58.002kDa and 56.401kDa,respectively,isoelectric point were 9.18 and 9.09.The homology of the two genes encoding the amino acid and the tartary buckwheat cinnamate-4-hydroxylase was 99% and 86% respectively,so it was named FtC4 H1 and FtC4 H2 temporarily.FtC4 H1 and FtC4 H2 were into two classes with orthologous proteins of other species.FtC4 H1 and FtC4 H2 were closed to the homologous proteins of Lettuce and Salvia miltiorrhiza ,and the amino acid sequence homology was 88% and 84%.[Conclusion]In this study,we analyzed the nucleic acid,amino acid sequence,protein structure and phylogenetic tree of two C4 H genes in tartary buckwheat,and provided the theo-retical basis for the subsequent extraction of phenylpropane and the utilization of buckwheat gene.关键词
苦荞/肉桂酸-4-羟基化酶/电子克隆/生物信息学分析Key words
Tartary buckwheat/Cinnamate-4-hydroxylase/Electronic cloning/Bioinformatics analysis分类
农业科技引用本文复制引用
刘荣华,王丽,孙朝霞,侯思宇,李红英..苦荞C4H基因的cDNA克隆及生物信息学分析[J].山西农业大学学报（自然科学版）,2017,37(11):767-773,7.基金项目
国家自然科学基金(NSFC:31301385) （NSFC:31301385）
山西省科技攻关项目(20150311007-1) （20150311007-1）
