• 생명과학회지
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• 제18권3호
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• pp.298-303
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• 2008

펩타이드 문고 기술을 이용하여 흑색종 세포주인 B16FI0에 결합하는 펩타이드 리간드를 검색하였다. 먼저 세포 내부로 들어간 파지들을 선택하는 방법으로 두 번 검색한 후 표면에 결합한 파지들 가운데 트랜스페린 단백질을 이용하여 트랜스페린 수용체에 결합한 파지들만을 선별적으로 용출시키는 방법으로 세 번 검색하였다. 다음으로 이 두 가지 방법을 통해 선별된 파지들에 표현된 펩타이드들을 Pseudomonas exotoxin의 전이 영역과 촉매 영역에 융합시킨 재조합 독소들을 만들었다. B16FI0 세포에 대한 각 재조합 독소의 활성을 측정하여 일곱 개의 클론을 선택한 후 염기서열을 분석하였다. 그 결과 그 가운데 한 클론에서 표현하는 펩타이드의 아미노산 서열이 사람의 트랜스페린과 유사한 서열을 갖는 것으로 확인되었다. 그 펩타이드를 화학적으로 합성한 후 항암제를 포함하는 리포솜에 붙여 실험한 결과 트랜스페린 수용체를 통해 치료물질을 전달할 수 있는 가능성을 지닌 것으로 평가되었다.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1518-1522
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• 2006

In the present study, it was observed how the phage-host system that is naturally reproduced in activated sludge is affected by the host inoculation. The system of Microlunatus phosphovorus and its phages was selected as the phage-host system native to an activated sludge system operated for 19 days under sequencing anaerobic-aerobic conditions with glutamate as the main carbon source. The phage-host system related to M. phosphovorus was monitored by plaque assay for the phages and by fluorescent in situ hybridization (FISH) for the bacterial host. In addition, the whole phage structure was also monitored by pulsed-field gel electrophoresis (PFGE). During the first 9 days, the phage-host system was more or less steady at approx. 9% (FISH/ DAPI) for M. phosphovorus and approx. 10,000 PFU/ml for its lytic phages. Microlunatus phosphovorus JCM9379 was inoculated into the activated sludge on day 10. Right after the inoculation, M. phosphovorus was approx. 24% (FISH/DAPI) whereas its lytic phages dropped down to approx. 500 PFU/ ml. After the host inoculation (within 9 days), however, the phage-host system eventually reverted to its original level in each population. On the other hand, the whole phage structure was not significantly changed by M. phosphovorus inoculation but stable throughout the process operation. Only the minor change that four phage groups gradually became abundant after the host inoculation was observed.

• 한국식품위생안전성학회지
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• 제35권6호
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• pp.602-606
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• 2020

본 연구는 다양한 항생제 내성을 갖는 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacininduced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM)에 대한 phage의 흡착 및 용균 특성을 평가하였다. PBST-10, PBST-13, PBST-32, PBST-35, P-22, P-22 B1 phages는 narrow host range를 보였다. 숙주인 STWT, STKCCM, STCIP에 대한 phage의 흡착률은 각각 47-85%, 58-95%, 61-93%였지만, STCCARM에 대한 phage의 흡착률은 14-36%의 낮은 수준을 보였다. STWT, STKCCM, STCIP, STCCARM에 대한 phage burst size는 각각 43-350, 37-530, 66-500, 24-500 plaque-forming unit(PFU)으로 다양하게 관찰되었다. P-22 B1을 제외한 모든 phage는 배양 초기에 STCIP숙주를 효과적으로 저해하였다. 이러한 결과는 항생제 내성균을 저해하기 위해 phage control system 개발에 유용한 정보로 활용될 것이다.

• The Plant Pathology Journal
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• 제34권1호
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• pp.59-64
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• 2018

Bacteriophages of Acidovorax citrulli, the causal agent of bacterial fruit blotch, were isolated from 39 watermelon, pumpkin, and cucumber leaf samples collected from various regions of Korea and tested against 18 A. citrulli strains. Among the six phages isolated, ACP17 forms the largest plaque, and exhibits the morphology of phages in the Myoviridae family with a head diameter of $100{\pm}5nm$ and tail length of $150{\pm}5nm$. ACP17 has eclipse and latent periods of $25{\pm}5min$ and $50{\pm}5min$, respectively, and a burst size of 120. The genome of ACP17 is 156,281 base pairs with a G + C content of 58.7%, 263 open reading frames, and 4 transfer RNA genes. Blast search and phylogenetic analysis of the major capsid protein showed that ACP17 has limited homology to two Stentrophomonas phages, suggesting that ACP17 is a new type of Myoviridae isolated from A. citrulli.

• 한국자기공명학회논문지
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• 제24권3호
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• pp.70-76
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• 2020

The CRISPR-Cas system provides an adaptive immunity for bacteria and archaea against invading phages or foreign plasmids. In the type II CRISPR-Cas system, a single effector protein Cas9 and a guide RNA form an RNA-guided endonuclease complex that can degrade DNA targets of foreign origin. To avoid the Cas9-mediated destruction, phages evolved anti-CRISPR (Acr) proteins that neutralize the host bacterial immunity by inactivating the CRISPR-Cas system. Here we report the backbone ¹H, ¹⁵N, and ¹³C resonance assignments of AcrIIA5 that inhibits the endonuclease activity of type II-A Streptococcus thermophilus Cas9 and also Streptococcus pyogenesis Cas9 using triple resonance nuclear magnetic resonance spectroscopy. The backbone chemical shifts of AcrIIA5 predict a disordered region at the N-terminus, followed by an αββββαβββ fold.

• 한국자기공명학회논문지
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• 제22권3호
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• pp.71-75
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• 2018

The CRISPR-Cas system is the adaptive immune system in bacteria and archaea against invading phages or foreign plasmids. In the type II CRISPR-Cas system, an endonuclease Cas9 cleaves DNA targets of phages as directed by guide RNA comprising crRNA and tracrRNA. To avoid targeting and destruction by Cas9, phages employ anti-CRISPR (Acr) proteins that act against host bacterial immunity by inactivating the CRISPR-Cas system. Here we report the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of AcrIIA4 that inhibits endonuclease activity of type II-A Listeria monocytogenes Cas9 and also Streptococcus pyogenesis Cas9 using triple resonance nuclear magnetic resonance spectroscopy. The secondary structures of AcrIIA4 predicted by the backbone chemical shifts show an ${\alpha}{\beta}{\beta}{\beta}{\alpha}{\alpha}$ fold, which is used to determine the solution structure.

• Journal of Microbiology and Biotechnology
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• 제30권10호
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• pp.1443-1457
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• 2020

The emergence and spread of antimicrobial resistance in pathogenic bacteria of fish and shellfish have caused serious concerns in the aquaculture industry, owing to the potential health risks to humans and animals. Among these bacteria, Aeromonas salmonicida, which is one of the most important primary pathogens in salmonids, is responsible for significant economic losses in the global aquaculture industry, especially in salmonid farming because of its severe infectivity and acquisition of antimicrobial resistance. Therefore, interest in the use of alternative approaches to prevent and control A. salmonicida infections has increased in recent years, and several applications of bacteriophages (phages) have provided promising results. For several decades, A. salmonicida and phages infecting this fish pathogen have been thoroughly investigated in various research areas including aquaculture. The general overview of phage usage to control bacterial diseases in aquaculture, including the general advantages of this strategy, has been clearly described in previous reviews. Therefore, this review specifically focuses on providing insights into the phages infecting A. salmonicida, from basic research to biotechnological application in aquaculture, as well as recent advances in the study of A. salmonicida.

• Journal of Microbiology
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• 제39권3호
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• pp.219-225
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• 2001

Phage typing is currently used for typing of Staphylococcus aureus strains beyond the species level in epidemiological studies. A total of 168 Staphylococcus aureus isolates from chicken meat and chicken by-products were phage-typed using the international bacteriophage set for typing Staphylococcus aureus of human origin. One hundred and forty-eight (88.79%) strains were phage-typeable (at least one phage produced 20 or more plaques of lysis). Lysis by phages of group Ⅲ was the mast frequent with 99 (58.93%) sensitive strains. This fact coincides with results of other authors. Twenty-nine different phage patterns were observed and three (95, 75/84 and 6/1030/W57) were most common. One hundred and thirty-two (89.19% of typeable strains) skewed these or indistinguishable (only one phage reaction difference) patterns. Twenty-six out of seventy chicken samples (37.14%) harboured more than one phage type of Staphylococcus aureus. This fact emphasizes the convenience of subtyping several Staphylococcus aureus isolates from the same sample in epidemiological studies. 80% of sausages and hamburgers contained the same Staphylococcus aureus phage types, which wore not found in any of the other food types. This fact suggests a cross contamination during the processing of these foods. Phages 6, 75, 84, 1030 and W57 skewed the greatest activity. None of the Staphylococcus aureus strains were sensitive to phages 47, 81 and 94.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1473-1480
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• 2016

This study focused on the variations in the non-coding sequences between ctxB and rstR of various CTX phages. The non-coding sequences of CTX-1 and CTX-cla are phage type-specific. The length of the non-coding region of CTX-1 and CTX-cla is 601 and 730 nucleotides, respectively. The non-coding sequence of CTX phage could be divided into three regions. There is a phage type-specific Variable region between two homologous Common regions (Common regions 1 and 2). The non-coding sequence of RS1 element is similar to CTX-1 except that Common region 1 is replaced by a short RS1-specific sequence. The non-coding sequences of CTX-2 and CTX-cla are homologous, indicating the non-coding sequence of CTX-2 is derived from CTX-cla. The non-coding region of CTX-O139 is similar to CTX-cla and CTX-2; however, it contains an extra phage type-specific sequence between Common region 2 and rstR. The variations in the non-coding sequences of CTX phages might be associated with the difference in the replication efficiency and the directionality in the integration into the V. cholerae chromosome.

• 한국식품과학회지
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• 제46권3호
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• pp.323-327
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• 2014

새싹채소로부터 분리된 E. faecium의 temperate phage 특성을 mitomycin C를 이용하여 E. faecium으로부터 D-19 phage와 F6 phage를 각각 분리하였다. 분리된 temperate phage는 형태학적 특성을 확인한 결과 모두 Siphoviridae에 속하는 것으로 나타났다. 그리고, 숙주 저해 범위는 55개의 숙주중에서 D-19 phage는 5주, F6 phage는 3주의 E. faecium만을 용균시킬 수 있는 것으로 확인하였다. 다양한 ethanol 농도에서의 안정성은 고농도에서도 매우 안정한 것으로 확인되었으며, pH의 안정성도 pH 4까지 안정한 것으로 나타났다. 본 연구를 통해 아직 연구가 많이 이루어지지 않은 E. faecium의 temperate phage는 host spectrum이 넓지 않은 것으로 나타났고 pH, 온도 등의 환경인자에 상당히 강한 안정성을 가지고 있는 것으로 나타났다.