霍乱弧菌噬菌体VP2基因组序列的测定与分析  

Complete Genome Sequence and Analysis of Vibrio Cholerae Phage VP2

作　　者：王多春[1] 汪敏 李燕萍[1] 董辉 刘中华[1] 刘延清[1] 祁国明[1] 高守一[1] 金维荣 阚飙[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所卫生部医学分子细菌学重点实验室,北京102206 [2]国家人类基因组南方研究中心,上海201203

出　　处：《病毒学报》2005年第1期60-64,共5页Chinese Journal of Virology

基　　金：国家重大基础研究发展规划(973计划)项目(G1999054102);国家自然科学基金(30271169)~~

摘　　要：测定并分析了霍乱弧菌噬菌体VP2基因组序列,为VP2生物学特性和功能研究提供分子遗传学基础。为此构建了VP2DNA随机文库,鸟枪法(shot gun)测定其全基因组序列。测序结果用软件Phrad Prap拼接成最小重叠群(contig),引物步移法测定contigs间的缝隙(gap)序列,拼接后获得VP2全基因组序列。利用生物信息学技术分析VP2基因组,最后对VP2和相关噬菌体做DNA聚合酶(DNApol)基因的进化树分析。结果:VP2属短尾噬菌体科,基因组全长39853bp,为环状双链DNA,G+C含量为50.56%,较高于霍乱弧菌测序菌株N16961基因组G+C含量;VP2的基因组有碱基使用偏性;预测和注释了45个开放读码框(ORF),分析了DNA复制基因、衣壳蛋白和DNA包装基因、侵染相关基因。DNApol进化树比较结果,VP2与链球菌噬菌体Cp 1和芽孢杆菌噬菌体GA 1分为一群。根据对VP2基因组序列的测定和分析预测了VP2的ORF,并分析了其中的功能基因,推测VP2在进化关系上属于噬菌体phi29样噬菌体。In order to sequence and analyze Vibrio cholerae phage VP2 genome,to provide molecular genetics data for further study of VP2 biology and functional genes,as well as VP2 origin and evolution,a VP2 DNA random library was constructed,shot-gun method was used for sequencing the whole genome.DNA sequences were assembled into contigs using Phrad-Prap Assembly System,the primer walking technique was used for sequencing the remaining fragments(gaps) and linking different contigs.Bioinformatics technique was used to analysis VP2 genome,phylogenetic trees of DNA polymerase (DNA pol) genes of VP2 and the related tailed phages were analyzed. The results showed that VP2 belongs to Podoviridae,the circular double-stranded DNA(ds DNA) VP2 genome consists of 39 853 bp,with overall G+C content of 50.56%,which was higher than the G+C content of N16961,a sequenced strain of Vibrio cholerae. Putative forty-five open reading frames(ORF) were annotated,homology analysis were used to assign possible functions for the genes,including, DNA replication,synthesis of capsid protein, DNA packaged genes, and infection-associated genes. The result of phylogenetic tree of DNA pol showed VP2 was clustered together with phage Cp-1 and GA-1. The evolutive analysis above suggests VP2 belongs to phi29-like phage.

关 键 词：霍乱弧菌 噬菌体 鸟枪法 VP2基因组 序列分析 DNA聚合酶 霍乱 

分 类 号：R516.5[医药卫生—内科学] R378.3[医药卫生—临床医学]