埃及伊蚊、冈比亚按蚊和致倦库蚊polyUBQ基因的电子克隆及其序列分析  

In silico Cloning and Sequence Analysis of polyUBQ Gene in Aedes aegypti,Anopheles gambiae and Culex quinquefasciatus

作　　者：强承魁[1] 周保亚[1] 凤舞剑[1] 赵虎[2] 苏新林[2] 王胜永[1] 

机构地区：[1]徐州生物工程职业技术学院农林工程系,徐州221006 [2]徐州生物工程职业技术学院教务处,徐州221006

出　　处：《生物技术通报》2011年第2期126-130,178,共6页Biotechnology Bulletin

基　　金：徐州生物工程职业技术学院院级科研课题(2010B07)

摘　　要：针对具有选择性蛋白质降解功能的泛素在调控昆虫生长发育过程中的重要作用,探讨埃及伊蚊、冈比亚按蚊和致倦库蚊基因组中polyUBQ基因的有关生物信息。采用电子克隆的方法钓取3种蚊虫基因组中polyUBQ基因序列,分析其特征、分子进化关系、遗传多态性和密码子偏爱性。结果显示,成功获取埃及伊蚊、冈比亚按蚊和致倦库蚊polyUBQ基因序列,命名为Aa-polyUBQ(GenBank登录号:AAGE02005963)、Ag-polyUBQ(ABKP02009650)和Cq-polyUBQ(AAWU01023041),分别编码1 065个、218个和533个氨基酸残基,各具14个、3个和7个泛素单体,Aa-polyUBQ、Ag-polyUBQ和Cq-polyUBQ蛋白二级结构主要元件是延伸带和无规则卷曲,Leu、Ile和Lys是构成3种蛋白的主要氨基酸,亚细胞主要定位于细胞质和细胞核,无前导肽、信号肽和跨膜结构域,除Ag-polyUBQ蛋白外均呈碱性;3种基因序列的同源性较高(83.8%-88.2%)且遗传距离较近(0.129-0.187);检出135个多态位点,共生成3个单倍型,单倍型多样性(Hd=1.000)、平均核苷酸差异数(K=96.000)、核苷酸多样性(P i=0.14679)、SS核苷酸多样性均值(P i(s)=0.57731)、NSS核苷酸多样性均值(P i(a)=0.01627)、密码子有效值(ENC=29.432-34.321<61)、偏爱指标(CB I=0.672-0.750>0)和2χ检验计算(2χ=0.977-1.092)显示3种polyUBQ基因间遗传多样性较丰富且呈现较强的密码子偏爱性。Aa-polyUBQ、Ag-polyUBQ和Cq-polyUBQ基因的成功电子克隆及丰富的遗传多态性和较强的密码子偏爱性可为进一步开展其分子进化机制、表达特性和昆虫病毒侵染关系等方面的试验研究提供基础资料。According to the important function of selective protein degradation of ubiquitin with regulating insect growth and development,relative bioinformation of polyUBQ gene in Aedes aegypti,Anopheles gambiae and Culex quinquefasciatus genome were investigated.The coding sequence of polyUBQ genes from three kinds of mosquitoes genome were found by in silico cloning method,then their sequence characteristics,molecular evolution,genetic polymorphism and code bias were analyzed.Results showed that polyUBQ gene sequences of Aedes aegypti,Anopheles gambiae and Culex quinquefasciatus named Aa-polyUBQ（GenBank Accession No.AAGE02005963）,Ag-polyUBQ（ABKP02009650）and Cq-polyUBQ（AAWU01023041）were obtained,which could code 1 065,218 and 533 amino acids with 14,3 and 7 repeats of ubiquitin coding unit,respectively.The main components of the predicated secondary structures of three proteins were extended strands and random coils,the principle amino acids of three proteins which consisted mainly of Leu,Ile and Lys.Subcells were mainly located in cytoplasm and cell nucleus,but leader peptide,signal peptide and transmembrane doamin were not found.Aa-polyUBQ and Cq-polyUBQ were characteristically basic accept for Ag-polyUBQ.Aa-polyUBQ,Ag-polyUBQ and Cq-polyUBQ gene sequences showed high identity（83.8%-88.2%）and short genetic distances（0.129-0.187）.During the genetic diversity analysis,total 153 polymorphic sites were detected from three polyUBQ gene sequences and 3 hapolotypes were sorted.Abundant genetic diversity and strong codon usage bias were found by haplotype diversity（Hd=1.000）,average number of nucleotide difference（K=96.000）,nucleotide diversity（Pi=0.14679）,nucleotide diversity of synonmyous sites（Pi（s）=0.57731）,nucleotide diversity of nonsynonmyous sites（Pi（a）=0.01627）,effective number of codons（ENC=29.432-34.32161）,codon bias index（CBI=0.672-0.7500）and scaled Chi-square（χ2=0.977-1.092）.The results of this investigation might provide some elements data for further st

关 键 词：埃及伊蚊 冈比亚按蚊 致倦库蚊 polyUBQ基因 电子克隆 

分 类 号：Q96[生物学—昆虫学] Q78