• Korean Journal of Fisheries and Aquatic Sciences
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• 제45권6호
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• pp.654-658
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• 2012

Vibrio alginolyticus, a marine fish and shellfish pathogen, has been found at a high frequency around the coastal areas of Korea. Vibrio alginolyticus was purified from various diseased fish, and a V. alginolyticus-specific bacteriophage was isolated from seawater obtained from fish farms located on the west coast of Korea. In a bacterial lysis experiment using the phage and antibiotics, tetracycline, $10^3$ cfu/ml of V. alginolyticus were completely lysed by both the phage and the antibiotic, suggesting that the purified phage in the study could be utilized as an alternative to antibiotics in the control of fish and shellfish diseases caused by V. alginolyticus.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• 제1권1호
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• pp.52-57
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• 2020

Bacterial symbionts are common across insects, including ants (Hymenoptera: Formicidae). Reproduction-manipulating endosymbionts, such as Wolbachia, Spiroplasma, Rickettsia, and Cardinium, are closely associated with many aspects of host-insect life. In addition, phage WO plays an essential role in the phenotypic effects of Wolbachia. Although endosymbionts are possible biological control agents, there is a lack of knowledge of their rate of infection of ants in Korea. We tested a range of Korean ant species for the presence of Wolbachia, Spiroplasma, Rickettsia, Cardinium, and phage WO by extracting DNA from the ants and using specific primer sets to test the status of infections. In addition, the mitochondrial cytochrome c oxidase I (COI) gene of the host ants was amplified to confirm the molecular identification and phylogenetic relationship between the hosts. We found that infection with Wolbachia (29.6% of species) is relatively common when compared with that of other endosymbionts. Only one species was infected with Spiroplasma. Infection with Rickettsia and Cardinium was not detected in the examined ants. Most Wolbachia in ants were infected with phage WO. Although the phenotypic effects of endosymbionts in ants are still unknown, this first survey of endosymbionts in Korea is the first step toward the use of reproduction-manipulating endosymbionts.

• BMB Reports
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• 제37권2호
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• pp.159-166
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• 2004

Angiostatin is a potent anti-angiogenic protein. To examine the angiostatin-interacting proteins, we used the display-cloning method with a T7 phage library presenting human cDNAs. The specific T7 phage clone that bound to the immobilized angiostatin was isolated, and a novel gene encoding the displayed polypeptide on the isolated T7 phage was identified. The displayed angiostatin-binding sequence was expressed in E. coli as a soluble protein and purified to homogeneity. This novel angiostatin-binding region interacted specifically to angiostatin with a dissociation constant of $3.4{\times}10^{-7}\;M$. A sequence analysis showed that the identified sequence was a part of the large ORF of 1,998 amino acids, whose function has not yet been characterized. A Northern analysis indicated that the gene containing the angiostatin-binding sequence was expressed differentially in the developmental stages or cell types.

• Bulletin of the Korean Chemical Society
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• 제31권12호
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• pp.3703-3706
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• 2010

Biomolecules specific to explosives can be exploited as chemical sensors. Peptides specific to immobilized dinitrotoluene (DNT) were identified using a phage display library. A derivative of DNT that contained an extended amine group, 4-(2,4-dinitrophenyl)butan-1-amine, was synthesized and immobilized using a self-assembled monolayer surface on gold. Filamentous M13 phages displaying random sequences of 12-mer peptides specific to the immobilized DNT-derivate were isolated from the M13 phage library by biopanning. A common peptide sequence was identified from the isolated phages and the synthesized peptides showed selective binding to DNT. When the peptide was immobilized on a quartz crystal microbalance (QCM) chip, it showed a binding signal to DNT, while toluene barely showed significant binding to the QCM chip. These results demonstrate that peptides screened by biopanning against immobilized DNT can be useful for quick and accurate detection of DNT.

• Bulletin of the Korean Chemical Society
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• 제34권2호
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• pp.460-464
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• 2013

Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with $K_D$ value of $6.0{\times}10^{-7}$ M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.

• Journal of Microbiology and Biotechnology
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• 제33권8호
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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.

• BMB Reports
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• 제39권1호
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• pp.55-60
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• 2006

We describe a phage display strategy, based on the differential resistance of proteins to denaturant-induced unfolding, that can be used to select protein variants with improved conformational stability. To test the efficiency of this strategy, wild-type and two stable variants of ${\alpha}_1$-antitrypsin (${\alpha}_1AT$) were fused to the gene III protein of M13 phage. These phages were incubated in unfolding solution containing denaturant (urea or guanidinium chloride), and then subjected to an unfavorable refolding procedure (dialysis at $37^{\circ}C$). Once the ${\alpha}_1AT$ moiety of the fusion protein had unfolded in the unfolding solution, in which the denaturant concentration was higher than the unfolding transition midpoint ($C_m$) of the ${\alpha}_1AT$ variant, around 20% of the phage retained binding affinity to anti-${\alpha}_1AT$ antibody due to a low refolding efficiency. Moreover, this affinity reduced to less than 5% when 10 mg/mL skimmed milk (a misfolding-promoting additive) was included during the unfolding/refolding procedure. In contrast, most binding affinity (>95%) remained if the ${\alpha}_1AT$ variant was stable enough to resist unfolding. Because this selection procedure does not affect the infectivity of M13, the method is expected to be generally applicable to the high-throughput screening of stable protein variants, when activity-based screening is not possible.

• Korean Journal of Food Science and Technology
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• 제25권1호
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• pp.46-51
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• 1993

The inactivating effect of iron(II)-ascorbate complex (Fe-Asc) on various phages was previously reported. This paper describes the molecular target in the phage virion attacked by Fe-Asc. The effect of Fe-Asc on protein was investigated with bovine serum albumin and the structural protein of phage J1. There were no differences in the SDS-polyacrylamide gel electrophoresis (patterns of these two proteins when either they were treated) with Fe-Asc or not. Also, there were no changes in the amino acid composition and ultraviolet spectrum of the proteins. The effects of Fe-Asc on DNA was investigated with pUC18 DNA, M13mpB DNA and ${\lambda}$ DNA as well as DNA from phage J1. Fe-Asc caused initially nicking of the subsequently form of pUC18 DNA to yield the open circular form and then subsequently the linear form. Strand breaks were also confirmed with M13mp8 DNA and ${\lambda}$ DNA as well as J1 DNA. The results indicate that the strand breaks in phage DNA could be responsible for the inactivation of phages by Fe-Asc.

• Microbiology and Biotechnology Letters
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• 제18권3호
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• pp.215-220
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• 1990

Prophage cured strain derivatives from Luctobacillirs araei YIT 9018 were isolated from thermoinducible mutant of the parent lysogenic strain. Two thermoinducible mutants were isolated from L. casei YIT 9018 strain treated with N-methyl-N'-nitro-N-nitrosoguanidine. Prophage cured strains were selected after heat induction of thermoinducible strains at $42^{\circ}C$ for 30 min in MRT medium containing anti- 4 FSV serum. The prophage cured strains, L. casei HYM 1213 and L. casei HYM 4024, could be used an indicator strain for temperate phage $\phi$ FSW. The growth, lactic acid producing ability and carbohydrates fermentation of L. casei HYM 1213 were similar to the parent L. cmei YIT 9018 strain, but A. casei HYM 4024 was not. One of the prophage cured strain, L. cmei HYM 1213, could be used industrially .to produce lactic acid beverages because this strah could not induce the virulent phage$\phi$FSV. The physiological characterization of L. casei HYM 1213 strain was similar to the parent L. casei YIT 9018 strain.

• Journal of Microbiology
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• 제44권3호
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• pp.363-368
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• 2006

In this study, native Byadyrhizobium strains were isolated from the host plant, Glycine max, harvested from fields in Madhya Pradesh, India, and were typed by Iytic rhizobiophages. Eight indigenous (Soy2, ASR011, ASR031, ASR032, MSR091, ISR050, ISR076 and ISR078) and two exotic strains (USDA123 and CB1809), all of which evidenced a distinct reaction with six phages, were employed in this study. The symbiotic interaction of these strains was studied initially using soybean cultivar JS335 in a sand culture in a controlled environment, and the efficiency was assessed based on the nodule number, nodule dry weight, plant dry weight, nitrogenase activity, and total accumulation of N per plant. Symbiotic effectiveness was found to be highest with the native phage-sensitive isolate ASR011, whereas it was at a minimum with the phage-resistant isolates, ISR050 and ISR078. Additionally, the effectiveness of these strains was evaluated using six soybean cultivars belonging to different maturity groups; namely, Brags, Lee, Pusa20, PK416, JS33S and NRC37. Analysis of variance data evidenced significant differences due to both symbionts, for the majority of the tested parameters. The CB1809, USDA123, and ASR011 strains evidenced relatively superior symbiotic effectiveness with soybean cultivars Brags, Lee and JS335. Strain ISR078 evidenced no significant responses with any of the cultivars. The ASR031 strain performed moderately well with all tested cultivars. The symbiotic response of all the strains was quite poor with cultivar PK416. Our studies showed that a significant relationship existed between the phage sensitivity and symbiotic efficiency of the bacterial strains with the host-cultivars.