茶小绿叶蝉代谢解毒基因CYP303A1的克隆及生物信息学分析

Cloning and Bioinformatics of Metabolic Detoxication Gene,CYP303A1,in Empoasca flavescens(Hemoptera:cicadas)

CYP303A1基因是细胞色素P450(简称P450)家族成员之一,它在昆虫药物代谢解毒过程中发挥重要作用。本研究通过RT-PCR方法克隆了茶小绿叶蝉Empoasca flavescens CYP303A1基因编码区ORF,并通过生物信息学软件和在线网站对该基因编码蛋白的理化性质、系统进化树、氨基酸多重序列比对、三级结构及功能域进行了分析。结果表明,CYP303A1基因含有1503bp开放阅读框,编码500个氨基酸,分子量为57.42kDa,理论等电点为8.56。系统进化树分析结果表明,CYP303A1基因与同翅目的烟粉虱Bemisia tabaci亲缘关系最近,与膜翅目的榕小蜂Ceratosolen solmsi亲缘关系最远。氨基酸系列比对结果表明,CYP303A1基因与其他物种昆虫CYP303A1的保守性较差。三级结构及功能域分析结果表明,该蛋白以β折叠为主,功能域由一信号肽和p450蛋白域(Pfam:p450)组成。该研究明确了茶小绿叶蝉CYP303A1的核苷酸序列及编码蛋白特征。

CYP303A1 gene belongs to the cytochrome P450 family.It plays an important role in metabolic detoxification of insecticides for insects.This study cloned the complete open reading frame（ORF）of CYP303A1 from Empoasca flavescens using RT-PCR.The physicochemical properties,phylogenetic tree,amino acid sequence,tertiary structure and functional domain of the gene were analyzed with the bioinformatics software and information from online sources.The results showed that the gene had a 1,503 bp ORF encoding 500 amino acids with a molecular weight of 57.42 kDa and a theoretical isoelectric point of 8.56.The phylogenetic tree analysis on the gene indicated that it had the highest genetic relationship with Bemisia tabaci（Homoptera）and the lowest with Ceratosolen solmsi（Hymenoptera）.The amino acid sequence of the gene poorly conserved with those of other insects,and the structure was primaryβ-sheet with its functional domain consisting of a signal peptide and pfam：p450.This study unveiled the nucleotide sequence and encoded amino acid characteristics of CYP303A1 from E.flavescens which would lead to the designing of a target insecticide for better control of E.flavescens on tea plants.