[Sequencing and analysis of Vibrio cholerae typing phage VP3 genome].
DNA sequence and the genome of phage VP3 (a typing phage of V. cholera) were analyzed. A random library of VP3 DNA was constructed by shot-gun library method. The VP3 genome sequence was assembled with contigs sequences, the gaps between different contigs were filled with sequencing data from primer walking. ORFs were predicted; Phylogeny of DNA polymerase sequences was analyzed to determine the class of VP3; The activity of putative promoter genes were analyzed using lacZ report system. VP3 genome is a 39504bp of circular double-stranded DNA. Twenty-seven out of forty-nine putative ORFs were annotated; twenty gene products were homologous with T7-like phages, including DNAP, DNA replicative protein, capsid, tail tubular, tail fiber protein, and DNA packaged protein. The activity of the putative promoter regions was confirmed through cloning those regions to LacZ-fuse plasmid pRS1274 and analysis of the expression of beta galactosidase. The complete genomic sequence of VP3 and phylogenetic tree analysis suggests VP3 is a member of T7 phage family.
Department of Microbiology, Medical School, Soochow University, Suzhou 215123, China. zhaoyingwei1993@sina.com
Ying-Wei
YW
Zhao
Duo-Chun
DC
Wang
Min
M
Wang
Yan-Ping
YP
Li
Biao
B
Kan
Hui
H
Dong
Zhong-Hua
ZH
Liu
Mei-Ying
MY
Yan
Wei
W
Li
Wei-Li
WL
Liang
Xue-Guang
XG
Zhang
Shou-Yi
SY
Gao
Wei-Rong
WR
Jin
D017123:Bacteriophage T7 / Q000145:classification / Q000235:genetics*; D001434:Bacteriophage Typing; D016679:Genome, Viral; D016366:Open Reading Frames; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D014734:Vibrio cholerae / Q000145:classification / Q000821:virology*