Rapid genetic characterization of a novel Enterobacteria phage and determination of its host recognizing genes.
We isolated a novel Enterobacteria phage IME08 from hospital sewage, then confirmed it was a double-stranded DNA phage by digesting its genetic material with DNase I, RNase A and several restriction endonucleases respectively. BLAST results of random fragments generated by a random PCR cloning method revealed that it belonged to T4-like virus. We subsequently determined the host recognizing genes (g37 and g38) sequence with a PCR-based "genome jumping" protocol based on highly conserved region at 5' terminus of g37 from four other T4-like Bacteriophages (T4, JS98, T2 and K3). These molecular biological methods enabled us to readily characterize the bacteriophage and efficiently determine the sequence of the genes of interest based on very limited conserved sequence information.
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
Huanhuan
H
Jiang
Sheng
S
Wang
Cun
C
Li
Dabin
D
Liu
Changming
C
Yu
Xiaoping
X
An
Zhiqiang
Z
Mi
Jiankui
J
Chen
Yigang
Y
Tong
D017122:Bacteriophage T4 / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000821:virology*; D016679:Genome, Viral / Q000235:genetics; D058507:Host Specificity / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods
D004279:DNA, Viral