• 미생물학회지
• /
• 제23권2호
• /
• pp.129-137
• /
• 1985

The RK-temperate phage which infected with Bacillus cereus was isolated and the characters were investigated. The induction of RK-temperate phage from host bacterium attained by ultraviolet light irradiation (15W, 30cm, 30-120sec) and mitomycin C treatment (0.2-2 ug/ml). The host range of RK-temperate phage was not revealed with lysogenic and related strains of B. cereus. But B. cereus(PS) 352 which obtained by N-nitrosoguanidine treatment (1,000{$\mu}g/ml)$ to phage infected with host bacteria was sensitive bacteria of RK-temperate phage. RK-temperate phage was stabilized at the condition of nutrient broth (pH 7-8), Tris-buffer (pH 7-8) and ammonium buffer (pH 8-9) and Sorensen's phosphate buffer (pH 6-7), but unstabilized at other salt solutions and pH range. Also, thermostability was to $45^{\circ}C$ but unstabilized at above $50^{\circ}C$. At RK-temperate phage, the measurment values of head, neck, mid tail and end tail were 59nm, $9{\times}16nm,\;10{\times}189nm,\;and\;10{\times}14nm$ respectively. The morphology of head was regular polyhedron, and the end tail was coneate form. On the one hand, the number of capsid protein layer of tail were consist of 4, 35, and 1 at neck, mid tail, and end tail, respectively. RK-temperate phage was identified with DNA phage and G+C contents were 38.63. The latent time of RK-temperate phage was 30 minutes and the burst size was 70-80. And the host bacteria was lysed in case of multi-infection, above moi 1.

• 미생물학회지
• /
• 제16권2호
• /
• pp.79-89
• /
• 1978

The RK-temperate phage which infected with Bacillus cereus was isolated and the characters were investigated. The induction of RK-temperate phage from host bacterium attained by ultraviolet light irradiation (15W, 30cm, 30-120sec) and mitomycin C treatment (0.2-2 ug/ml). The host range of RK-temperate phage was not revealed with lysogenic and related strains of B. cereus. But B. cereus(PS) 352 which obtained by N-nitrosoguanidine treatment(1,000.$\mu$g/ml) to phage infected with host bacteria was sensitive bacteria of RK-temperate phage. RK-temperate phage was stabilized at the condition of nutrient broth (pH 7-8), Tris-buffer (pH 7-8) and ammonium buffer (pH 8-9) and Sorensen's phosphate buffer (pH 6-7), but unstabilized at other salt solutions and pH range. Also, thermostability was to 45.deg.C but unstabilized at above 50.deg.C. At RK-temperate phage, the measurment values of head, neck, mid tail and end tail were 59nm, 9*16nm, 10*189nm, and 10*14nm respectively. The morphology of head was regular polyhedron, and the end tail was coneate form. On the one hand, the number of capsid protein layer of tail were consist of 4, 35, and 1 at neck, mid tail, and end tail, respectively. RK-temperate phage was identified with DNA phage and G+C contents were 38.63. The latent time of RK-temperate phage was 30 minutes and the burst size was 70-80. And the host bacteria was lysed in case of multi-infection, above moi 1.

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

• 한국어병학회지
• /
• 제11권2호
• /
• pp.137-141
• /
• 1998

TSB 해수배지와 숙주로서 L. garvieae No. 44를 배양조건으로 사용한 경우, L. garvieae 111균주중 용원화되었다고 추정되어진 96균주의 cells에서 temperate Phage가 효과적으로 분리되었다. 하지만 동일한 배양조건에서 보통의 TSB 배지를 사용한 경우에서는 temperate phage는 전혀 나타나지 않았다. 이 temperate phages는 TSB 해수배지와 ultraviolet irradiation를 병용한 경우 역시 효과적으로 분리되었다. 분리된 모든 temperate Phages는 기존의 phage(PLgY, PLgW, PLgS) 의 lytic nature와는 달리 오직 L.garvieae No. 44만을 lysis하였다. 배양후 약 1시간후 phage가 나타났으며 12시간 후 virulent phage의 최고농도($10^{10}$ PFU/ml) 보다 훨씬 낮은 농도인 $10^6$ PFU/ml까지 증가하였다.

• Journal of Microbiology and Biotechnology
• /
• 제22권5호
• /
• pp.649-653
• /
• 2012

A temperate phage was isolated from emetic Bacillus cereus NCTC 11143 by mitomycin C and characterized by transmission electron microscopy and DNA and protein analyses. Whole genome sequencing of Bacillus phage 11143 was performed by GS-FLX. The phage has a dsDNA genome of 39,077 bp and a 35% G+C content. Bioinformatic analysis of the phage genome revealed 49 putative ORFs involved in replication, morphogenesis, DNA packaging, lysogeny, and host lysis. Bacillus phage 11143 could be classified as a member of the Siphoviridae family by morphology and genome structure. Genomic comparisons at the DNA and protein levels revealed homologous genetic modules with patterns and morphogenesis proteins similar to those of other Bacillus phages. Thus, Bacillus phages might have a mosaic genetic relationship.

• 한국미생물·생명공학회지
• /
• 제18권3호
• /
• pp.215-220
• /
• 1990

L.casei YIT 9018균주에 N-methyl-N'-nitro-N-nitrosoguanidine을 처리하여 thermoinducible mutants를 분리하였고, 이 균주를 $42^{\circ}C$에서 30분 동안 열처리하여 prophage가 cured된 균주 L.casei HYM 1213과 L.casei HYM 4024를 분리하였다. Prophage cured strain L.case HYM 1213과 L.casei HYM 4024는 temperate phage $\phi$Fsw에 대해 지시균으로서 능력을 가지고 있었으며, 어떤 온도에서도 temperate phage $\phi$FSW가 검출되지 않았다. 이들의 생리적 특성을 모균주인 L.caseiYIT 9018과 비교한 결과 L.casei HYM 1213 균주는 균의 증식, pH변화, 산생성능력, 당 발효능력에서 모균주와 유사한 것으로 나타났으나, L.casei HYM 4024균주는 pH변화와 산생성능력이 모균주에 비해 미약한 것으로 나타났다. Plasmid DNA는 두 균주 모두 모균주와 동일한 size를 보여주여, prophage가 cured되었어도 plasmid DNA에는 손실을 가져오지 않았음을 알 수 있었다.

• Journal of Microbiology and Biotechnology
• /
• 제26권2호
• /
• pp.263-269
• /
• 2016

Temperate phages have been suggested to carry virulence factors and other lysogenic conversion genes that play important roles in pathogenicity. In this study, phage TEM123 in wild-type Staphylococcus aureus from food sources was analyzed with respect to its morphology, genome sequence, and antibiotic resistance conversion ability. Phage TEM123 from a mitomycin C-induced lysate of S. aureus was isolated from foods. Morphological analysis under a transmission electron microscope revealed that it belonged to the family Siphoviridae. The genome of phage TEM123 consisted of a double-stranded DNA of 43,786 bp with a G+C content of 34.06%. A bioinformatics analysis of the phage genome identified 43 putative open reading frames (ORFs). ORF1 encoded a protein that was nearly identical to the metallo-β-lactamase enzymes that degrade β-lactam antibiotics. After transduction to S. aureus with phage TEM123, the metallo-β-lactamase gene was confirmed in the transductant by PCR and sequencing analyses. In a β-lactam antibiotic susceptibility test, the transductant was more highly resistant to β-lactam antibiotics than S. aureus S133. Phage TEM123 might play a role in the transfer of β-lactam antibiotic resistance determinants in S. aureus. Therefore, we suggest that the prophage of S. aureus with its exotoxin is a risk factor for food safety in the food chain through lateral gene transfer.

• 한국미생물생명공학회:학술대회논문집
• /
• 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
• /
• pp.250-256
• /
• 2004

• BMB Reports
• /
• 제40권5호
• /
• pp.740-748
• /
• 2007

To gain insight into the structure and function of repressor proteins of bacteriophages of gram-positive bacteria, repressor of temperate Staphylococcus aureus phage ${\phi}11$ was undertaken as a model system here and purified as an N-terminal histidine-tagged variant (His-CI) by affinity chromatography. A ~19 kDa protein copurified with intact His-CI (~ 30 kDa) at low level was resulted most possibly due to partial cleavage at its Ala-Gly site. At ~10 nM and higher concentrations, His-CI forms significant amount of dimers in solution. There are two repressor binding sites in ${\phi}11$ cI-cro intergenic region and binding to two sites occurs possibly by a cooperative manner. Two sites dissected by HincII digestion were designated operators $O_L$ and $O_R$, respectively. Equilibrium binding studies indicate that His-CI binds to $O_R$ with a little more strongly than $O_L$ and binding species is probably dimeric in nature. Interestingly His-CI binding affinity reduces drastically at elevated temperatures ($32-42^{\circ}C$). Both $O_L$ and $O_R$ harbor a nearly identical inverted repeat and studies show that ${\phi}11$ repressor binds to each repeat efficiently. Additional analyses indicate that ${\phi}11$ repressor, like $\lambda$ repressor, harbors an N-terminal domain and a C-terminal domain which are separated by a hinge region. Secondary structure of ${\phi}11$ CI even nearly resembles to that of $\lambda$ phage repressor though they differ at sequence level. The putative N-terminal HTH (helix-turn-helix) motif of ${\phi}11$ repressor belongs to the HTH -XRE-family of proteins and shows significant identity to the HTH motifs of some proteins of evolutionary distant organisms but not to HTH motifs of most S. aureus phage repressors.

• BMB Reports
• /
• 제29권5호
• /
• pp.448-454
• /
• 1996

Bacteriophage ${\phi}FC1$ is a temperate phage which was identified as a prophage in the Enterococcus faecalis KBL703 chromosome. Phage ${\phi}FC1$ integrates into the host chromosome by site-specific recombination. The phage attachment site P (attP) was localized within the 0.65-kb XhoI-HindIII fragment and the nucleotide sequence of the region was determined. An open reading frame (mj1) which adjoined the phage attachment site encoded a deduced protein related to the site-specific recombinase family. The organization of this region was comparable to other site-specific recombination systems. The molecular weight of the expressed MJ1 in E. coli was in good agreement with the predicted 53,537 Da of the mj1 gene product. Elucidation of the phage-specific integration process in this study would provide useful genetic tools such as a chromosomal integration system.