• Korean Journal of Microbiology
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• v.32 no.1
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• pp.60-64
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• 1994

Pseudomonas syringae pv. tabaci produces tabtoxin and causes wildfire disease on tabacco and bean plants. In this study, bacteriophage of P. syringae pv. tabaci were isolated from sewage by top agar overlay method, and physiological and genetical characteristics of the phage were investigated. Plaques of isolated phage were turbid and ranged in size from 1 to 2 mm. The stability range of pH was between 6.0 and 9.0, and stability of temperature was up to 30${\circ}C$ and inactivated at 70${\circ}C$. The adsorption rate of phage was about 85% for 30min. The latent period and mean burst size as dertermined in one step growth experiments were 3 hrs and 200 PFU/bacterium, respectively. Genomic material of isolated phage was dsDNA of which size was about 30kb.

• Korean Journal of Food Science and Technology
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• v.10 no.2
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• pp.136-142
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• 1978

The young cells of Clostridiunm saccaroperbutylacetonicum were rapidly autolysed by exposing them to the hypertonic solution of sucrose(0.3-0.6M) without any other supplement to decompose the rigid cell wall. The cells were converted into the spherical cells by lysis. The spherical cells had following properties: (1) they were absent in the cell wall and osmotically fragile. (2) they were stabilized in the existence of 0.4M sucrose and 5mM $MgSO_4$ (3) they were resistant against adsorption of phage particles. (4) they allowed infection of the isolated phage DNA and produced progeny phage particles. (5) they were able to biosynthesize their macromolecules for a few hours according to a balanced manner of biosynthesis. (6) they were able to produce the bacteriocin particles by mitomycin C treatment. (7) they were unable to multiply. These results were all in the level of typical properties of bacterial protoplasts. It was apparent that the spherical cells formed by lysis occcurring by treatment with hypertonic sucrose were protoplasts.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.510-518
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• 2021

Bacteriophages are considered excellent sensing elements for platforms detecting bacteria. However, their lytic cycle has restricted their efficacy. Here, we used the minor coat protein pIII domain (N1N2) of phage CTXφ to construct a novel surface plasmon resonance (SPR) biosensor that could detect Vibrio cholerae. N1N2 harboring the domains required for phage adsorption and entry was obtained from Escherichia coli using recombinant protein expression and purification. SDS-PAGE revealed an approximate size of 30 kDa for N1N2. Dot blot and transmission electron microscopy analyses revealed that the protein bound to the host V. cholerae but not to non-host E. coli K-12 cells. Next, we used amine-coupling to develop a novel recombinant N1N2 (rN1N2)-functionalized SPR biosensor by immobilizing rN1N2 proteins on gold substrates and using SPR to monitor the binding kinetics of the proteins with target bacteria. We observed rapid detection of V. cholerae in the range of approximately 10³ to 10⁹ CFU/ml but not of E. coli at any tested concentration, thereby confirming that the biosensor exhibited differential recognition and binding. The results indicate that the novel biosensor can rapidly monitor a target pathogenic microorganism in the environment and is very useful for monitoring food safety and facilitating early disease prevention.

• Journal of Life Science
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• v.28 no.1
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• pp.78-82
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• 2018

Detection of pathogenic microorganisms takes several days by conventional methods. It is necessary to assess microorganisms in a timely manner to reduce the risk of spreading infection. For this purpose, bacteriophages are chosen for use as a biosensing tool due to their host specificity, wide abundance, and safety. However, their lytic cycle limits their efficacy as biosensors. Phage proteins involved in binding to bacteria could be a robust alternative in resolving this drawback. Here, a fragment of tail protein J (residues 784 to 1,132) of phage lambda fused with 6X His-tag (6HN-J) at its N-terminus was cloned, overexpressed, purified, and characterized for its binding with microorganisms. The purified protein demonstrated a size of about 38 kDa in sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and bound with anti-His monoclonal antibodies. It bound specifically to Escherichia coli K-12, and not Salmonella typhimurium, Bacillus subtilis, or Pseudomonas aeruginosa in dot blotting. Binding of the protein to E. coli K-12 inhibited about 50% of the in vivo adsorption of the phage lambda to host cells at a concentration of $1{\mu}g/ml$ 6HN-J protein and almost 100% at $25{\mu}g/ml$ 6HN-J. The results suggest that a fusion viral protein could be utilized as a biosensing element (e.g., protein chips) for detecting microorganisms in real time.