[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.
Zhao
Ying-Wei
YW
Department of Microbiology, Medical School, Soochow University, Suzhou 215123, China. zhaoyingwei1993@sina.com
Wang
Duo-Chun
DC
Wang
Min
M
Li
Yan-Ping
YP
Kan
Biao
B
Dong
Hui
H
Liu
Zhong-Hua
ZH
Yan
Mei-Ying
MY
Li
Wei
W
Liang
Wei-Li
WL
Zhang
Xue-Guang
XG
Gao
Shou-Yi
SY
Jin
Wei-Rong
WR
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*