中华按蚊血红素过氧化物酶家族基因的全基因组鉴定、特征和进化  

Genome-wide identification,characterization and evolutionary analysis of genes of the heme peroxidase family in Anopheles sinensis(Diptera:Culicidae)

作　　者：尹华春 张莉娟 陈斌[1] 

机构地区：[1]重庆师范大学昆虫与分子生物学研究所,媒介昆虫重庆市重点实验室,重庆401331

出　　处：《昆虫学报》2018年第1期36-47,共12页Acta Entomologica Sinica

基　　金：“两江学者”计划专项经费;国家自然科学基金项目(31672363,31372265);国际原子能机构CRP项目(18268);国家科技基础性工作专项重点项目(2015FY210300);广西科技重大专项(桂科AA17129002)

摘　　要：【目的】在全基因组上鉴定中华按蚊Anopheles sinensis血红素过氧化物酶(heme peroxidase,HPX)家族基因,并预测其基本特征,探究双翅目中5种代表性昆虫HPX基因的系统发育关系和进化。【方法】以NCBI数据库中黑腹果蝇Drosophila melanogaster和家蚕Bombyx mori等昆虫HPX基因编码的氨基酸序列为询问序列,通过本地Blast搜索鉴定中华按蚊基因组上HPX家族基因,并鉴定了冈比亚按蚊Anopheles gambiae、致倦库蚊Aedes aegypti和埃及伊蚊Culex quinquefasciatus基因组上的HPX基因;基于冈比亚按蚊HPX基因的命名系统,对中华按蚊HPX基因进行命名;运用生物信息学方法预测了中华按蚊HPX基因的特征,包括基因的结构及在scaffold的定位,氨基酸的替换率和保守结构域,蛋白质3D结构等,通过与冈比亚按蚊共线性分析定位了中华按蚊HPX基因在染色体上的位置;基于HPX核苷酸序列,采用PAUP4.0和MEGA6.0软件利用最大相似法构建了5个双翅目代表性种HPX基因的系统发生树。【结果】中华按蚊基因组共有20个HPX基因,冈比亚按蚊、埃及伊蚊和致倦库蚊基因组分别有18,14和12个HPX基因。这4种蚊虫的HPX蛋白都被分类进入Peroxinectin,Peroxidasin,DBLPX和DUOX 4个亚家族,其氨基酸的分子量介于61.6~186.6 k D之间(除As HPX8为29.6 k D)。中华按蚊20个HPX基因共具有98个外显子,75个内含子,其外显子与内含子分布模式在基因间差异较大。中华按蚊HPX基因被定位到10个scaffold上,对应到冈比亚按蚊的2R,3R,2L,3L和X染色体。这些中华按蚊HPX基因编码的氨基酸序列(除DUOX)都具有1个血红素结合位点和5个Ca^(2+)结合位点,在N端和C端各具有2个半胍氨酸位点并界定了两个二硫键。中华按蚊与冈比亚按蚊同源基因对的ω值都小于1,说明HPX基因在进化过程中没有受到明显的环境选择压力。系统发育关系研究表明,5种双翅目昆虫的HPX基因可以分为16个组,其中11个组在进�【Aim】 To identify genes of the heme peroxidase （HPX） family of Anopheles sinensis at the wholegenome level, to predict the basic features of these HPX members, and to explore the phylogenetics and evolution of the HPX members of five representativedipterans. 【Methods】 The amino acid sequences encoded by HPX genes in Drosophila melanogaster, Bombyx mori and other insects were downloaded from NCBI databases and used as queries to search for the HPX genes in An. sinensis genome using the local Blast program. The HPX genes in Anopheles gambiae, Aedes aegypti and Culex quinquefasciatus on the whole genome were also identified by using the same method. The HPX genes identified in An. sinensis were named following the nomenclature system established for An. gambiae HPX genes. The characteristics of these HPX genes in An. sinensis, including the structure and scaffold location of these genes, the substitution rate and conserved domain of their amino acids, and the 3D structure of their proteins, were predicted using bioinformatics analysis. These An. sinensis HPX genes were also located on chromosome through the synteny analysis with An. gambiae HPX genes. The phylogenetic relationships of the HPX genes of five representative species of Diptera were constructed based on nucleotide sequences using maximum likelihood method with PAUP 4.0 and MEGA 6.0. 【Results】 The An. sinensis genome contains 20 HPX genes, while those of An. gambiae, Ae. aegypti and Cx. quinquefasciatus contain 18, 14 and 12 HPX genes, respectively. The HPX proteins of the four mosquito species were all classified into Peroxinectin, Peroxidasin, DBLPX and DUOX subfamilies, with the molecular weights of their amino acids ranging from 61.6-186.6 kD, except AsHPX8 （29.6 kD）. The 20 HPX genes of An. sinensis contain 98 exons and 75 introns, and the distribution patterns of their exons and introns are quite diverse among different genes. These HPX genes of An. sinensis are mapped on 10 scaffolds, and syntenied to the chromosomes 2R, 3R,

关 键 词：中华按蚊 HPX家族 全基因组鉴定 特征 系统发育 进化 

分 类 号：Q969[生物学—昆虫学]