• Korean Journal of Microbiology
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• v.39 no.2
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• pp.118-122
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• 2003

To develop vector plasmid for the bacteriophage P2-P4 system which is a useful experimental tool for the study of viral capsid assembly, we constructed a new P4-derived vector plasmid starting from P4 ash8 sid71 With recombinant DNA technology, a portion of P4 genome was deleted and tetracyclin resistance gene (terR) was introduced into P4 genome to give P4 selectivity. Resulting P4 ash8(sid71) terR was 12.09 kb long and could be converted to a viable bacteriophage with P2 infection. The burst size of induced bacteriophage form of P4 ash8(sid71) terR was determined. The CsCl buoyant equilibrium density gradient experiment of new P4 derivative suggested the upper limit of packaging capacity in P2-size head.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.77-78
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• 2018

Staphylococcus aureus is a Gram-positive and a round-shaped bacterium of Firmicutes phylum, and is a common cause of skin infections, respiratory infections, and food poisoning. Bacteriophages infecting S. aureus can be an effective treatment for S. aureus infections. Here, the draft genomic sequence is announced for a lytic bacteriophage SA7 infecting S. aureus isolates. The bacteriophage SA7 was isolated from a sewage water sample near a livestock farm in Chungcheongnam-do, South Korea. SA7 has a genome of 34,730 bp and 34.1% G + C content. The genome has 53 protein-coding genes, 23 of which have predicted functions from BLASTp analysis, leaving the others conserved proteins with unknown function.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.194-198
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• 2015

Enterococcus faecalis is a Gram-positive and facultative anaerobic bacterium that causes many hospital-acquired infections. Novel E. faecalis specific bacteriophage (phage) ECP3 that had been isolated from thirty-four environmental samples and characterized phenotypically and genotypically. ECP3 phage belongs to the family Myoviridae with contractile tail and lysed E. faecalis specifically but other bacteria including Enterococcus faecium. The genome was double-stranded linear DNA and its size was 145,518 bp comprising of 220 open reading frames. The GC content was 35.9%. The genome sequence showed 97% identity in 90% coverage region with Myoviridae phage PhiEF24C. ECP3 is the first E. faecalis-specific Myoviridae phage isolated in Korea which can be a promising antimicrobial agent against E. faecalis infections.

• Korean Journal of Microbiology
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• v.35 no.4
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• pp.266-269
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• 1999

Bacteriophage E3 grows very rapidly and forms a large size plaque with a diameter of 1 cm. The promoter controlling the expression of the gene encoding the major capsid protein is thought to be most efficient. To find out this promoter, this gene was mapped in the genome according to the following procedure. The major capsid protein was purified from phage particle and the N-terminal amino acid sequence was revealed. Based on this sequence,a degernerate oligonucleotide probe was designed and used for screening of the genomic DNA fragments. From the DNA sequence of the selected clone, the gene encoding the major capsid protein was mapped at 70% of E3 genome. The expression of this gene was not sensitive to rifampicin which indicated the presence of E3's own RNA polymerase.

• Journal of Life Science
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• v.13 no.4
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• pp.505-510
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• 2003

The extremely halophilic archaebacteriurn Haloarcular sp. EH-1 was isolated from solar salts. Halophages found in Haloarcular sp. EH-1 were isolated from fermented anchovy sauce. Halophages were isolated from fermented anchovy sauce using Haloarcular sp. EH-1 as a host bacterium. The isolated halophage produced 0.5∼l.0 mm in diameter clear plaques. The halophage consists of an symmetrical head, measuring 68 nm in diameter, and a contractile tail, 100 nm long and base plates were observed. Total size of phage DNA genome obtained 20 Kbp and its sequence homology was 52.87% with H. Salinarium.

• Microbiology and Biotechnology Letters
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• v.51 no.4
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• pp.562-565
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• 2023

Escherichia coli-specific phage, KFS-EC1, was isolated and purified from a slaughterhouse. The complete genome of the phage was obtained using Illumina MiSeq platforms. Its assembled genome consisted of a single chromosome of 164,715 bp with a GC content of 40.5%. The phage genome contained 170 hypothetical and 101 functional ORFs, and exhibited orthologous average nucleotide identity values of >95% with other E. coli phages belonging to the family Straboviridae. Additionally, phylogenetic analysis revealed that KFS-EC1 was finally classified into the family Straboviridae of the genus Caudoviricetes. The genome has been deposited in GenBank under the accession number NC_055757.1.

• BMB Reports
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• v.35 no.6
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• pp.637-641
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• 2002

Only one natural promoter that interacts with bacteriophage K11 RNA polymerase has so far been identified. To identify more, in the present study restriction fragments of the phage genome were individually assayed for transcription activity in vitro. The K11 genome was digested with two 4-bp-recognizing restriction enzymes, and the fragments cloned in pUC119 were assayed with purified K11 RNA polymerase. Eight K11 promoter-bearing fragments were isolated and sequenced. We report that the nine K11 promoter sequences (including the one previously identified) were highly homologous from -17 to +4, relative to the initiation site at +1. Interestingly, five had -10G and -8A, while the other four had -10A and -8C. The consensus sequences with the natural -10G/-8A and -10A/-8C, and their variants with -10G/-8C and -10A/-8A, showed nearly equal transcription activity, suggesting residues at -10 and -8 do not regulate promoter activity. Using hybridization methods, physical positions of the cloned promoter-bearing sequences were mapped on SalI-and KpnI-restriction maps of the K11 genome. The flanking sequences of six cloned K11 promoters were found to be orthologous with T7 or T3 genomic sequences.

• BMB Reports
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• v.48 no.1
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• pp.6-12
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• 2015

Various biological molecules naturally existing in diversified species including fungi, bacteria, and bacteriophage have functionalities for DNA binding and processing. The biological molecules have been recently actively engineered for use in customized genome editing of mammalian cells as the molecule-encoding DNA sequence information and the underlying mechanisms how the molecules work are unveiled. Excitingly, multiple novel methods based on the newly constructed artificial molecular tools have enabled modifications of specific endogenous genetic elements in the genome context at efficiencies that are much higher than that of the conventional homologous recombination based methods. This minireview introduces the most recently spotlighted molecular genome engineering tools with their key features and ongoing modifications for better performance. Such ongoing efforts have mainly focused on the removal of the inherent DNA sequence recognition rigidity from the original molecular platforms, the addition of newly tailored targeting functions into the engineered molecules, and the enhancement of their targeting specificity. Effective targeted genome engineering of mammalian cells will enable not only sophisticated genetic studies in the context of the genome, but also widely-applicable universal therapeutics based on the pinpointing and correction of the disease-causing genetic elements within the genome in the near future.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2151-2155
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• 2017

Bacteriophages have gained substantial attention as biocontrol and biorecognition agents, substituting antibodies. In this study, a Salmonella Enteritidis-specific bacteriophage, KFS-SE1, was isolated, identified, and characterized. This Siphoviridae phage infects S. Enteritidis with high specificity. This phage is highly stable under various pH (5-11), temperature ($4-60^{\circ}C$), and organic solvent conditions. The KFS-SE1 genome consisted of 59,715 bp with 73 predicted open reading frames and 57.14% GC content; it had a complete set of genes required for phage reconstruction. Comparative phylogenetic analysis of KFS-SE1 revealed that it was very similar to the other Salmonella phages in the Siphoviridae family. These characteristics suggest that KFS-SE1 with its high specificity and host lysis activity toward S. Enteritidis may have various potential applications.

• Journal of Microbiology and Biotechnology
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• v.33 no.8
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• pp.1050-1056
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• 2023

Weizmannia coagulans (formerly Bacillus coagulans) is Gram-positive, and spore-forming bacteria causing food spoilage, especially in acidic canned food products. To control W. coagulans, we isolated a bacteriophage Youna2 from a sewage sludge sample. Morphological analysis revealed that phage Youna2 belongs to the Siphoviridae family with a non-contractile and flexible tail. Youna2 has 52,903 bp double-stranded DNA containing 61 open reading frames. There are no lysogeny-related genes, suggesting that Youna2 is a virulent phage. plyYouna2, a putative endolysin gene was identified in the genome of Youna2 and predicted to be composed of a N-acetylmuramoyl-L-alanine amidase domain (PF01520) at the N-terminus and unknown function DUF5776 domain (PF19087) at the C-terminus. While phage Youna2 has a narrow host range, infecting only certain strains of W. coagulans, PlyYouna2 exhibited a broad antimicrobial spectrum beyond the Bacillus genus. Interestingly, PlyYouna2 can lyse Gram-negative bacteria such as Escherichia coli, Yersinia enterocolitica, Pseudomonas putida and Cronobacter sakazakii without other additives to destabilize bacterial outer membrane. To the best of our knowledge, Youna2 is the first W. coagulans-infecting phage and we speculate its endolysin PlyYouna2 can provide the basis for the development of a novel biocontrol agent against various foodborne pathogens.