樟叶越桔CYP450基因片段克隆与家族分析

Cloning and Family Analysis of CYP450 Gene Fragment from Vaccinium dunalianum Wight

广泛存在于生物体内的CYP450具有参与多种内源物质的新陈代谢和解除外源化学物质毒害作用的功能。本研究基于RNA-Seq数据,应用RT-PCR技术首次从樟叶越桔中克隆获得21个CYP450基因cDNA片段。应用生物信息学技术对21个樟叶越桔CYP450基因进行相似性分析、功能注释、家族分类以及聚类分析,除2个基因的家族分类不明确外,其余所有基因被分属于10个不同的CYP450家族中,分别为:CYP71、CYP72、CYP76、CYP81、CYP82、CYP86、CYP94、CYP97、CYP734和CYP735。推测有10个基因参与脂肪族、芳香族碳的羟基化,4个基因参与植物激素细胞分裂素的合成与降解,3个基因参与脂肪酸的羟基化,2个基因参与萜类化合物的合成。而KEGG分析结果显示2个基因家族未知的樟叶越桔CYP450基因可能参与了K00327途径。21个樟叶越桔CYP450基因能成功地与24个拟南芥CYP450基因聚类到一起。值得关注的是有2个樟叶越桔CYP450基因CL75.Contig12A(9)和Unigene225702A(14)被归类到拟南芥所特有的CYP82家族中,分别属于CYP82D和CYP82G亚家族,这为研究CYP82基因家族新物种来源提供了参考。本研究结果为认识樟叶越桔细胞色素CYP450基因和家族分类提供了信息,为樟叶越桔CYP450功能研究提供了参考依据。

Cytochrome P450(CYP450), which existing in organisms widely, is capable of participating in the metabolism of various endogenous substances and relieving the toxic effects of exogenous chemicals. In this study,based on RNA-Seq data, twenty-one c DNA fragments of CYP450 genes were cloned from Vaccinium dunalianum Wight for the first time by RT-PCR. Bioinformatics was used to analyze the similarity, functional annotation, family classification and cluster analysis of the 21 CYP450 genes in V. dunalianum. Except that the family classification of the two genes was not clear, all other genes were classified into 10 different CYP450 families: CYP71, CYP72,CYP76, CYP81, CYP82, CYP86, CYP94, CYP97, CYP734 and CYP735. It was speculated that 10 genes were involved in the hydroxylation of aliphatic and aromatic carbon, 4 genes were involved in the synthesis and degradation of plant hormone cytokinin, 3 genes were involved in the hydroxylation of fatty acids, and 2 genes were involved in the synthesis of terpenoids. The results of KEGG analysis showed that the 2 genes with unknown family classification could be involved in the K00327 pathway. The 21 CYP450 genes in V. dunalianum were successfully clustered with 24 CYP450 genes from Arabidopsis thaliana in the phylogenetic tree. It was worth noting that the two CYP450 genes CL75.Contig12 A(9) and Unigene225702 A(14) in V. dunalianum were classified into the CYP82 family unique to Arabidopsis thaliana, belonging to the CYP82 D and CYP82 G subfamilies, respectively, which provided a reference for the study of the new species source of the CYP82 family.The results of this study provided information for understanding the CYP450 genes and their family classification and a reference for the study of CYP450 function in V. dunalianum.