• 제목/요약/키워드: Selective Enrichment

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Development of a Selective Medium for the Fungal Pathogen Cylindrocarpon destructans Using Radicicol

  • Kang, Yunhee;Lee, Seung-Ho;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.432-436
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    • 2014
  • The soil-borne ascomycete fungus Cylindrocarpon destructans causes ginseng root rot disease and produces various secondary metabolites such as brefeldin A and radicicol. The slow growth of this fungus compared with other plant pathogenic and saprophytic fungi in soil disturbs isolation of this fungus from soil and infected ginseng. In this study, we developed a selective medium for C. destructans using radicicol produced by this fungus. Supplementing 50 mg/L of radicicol to medium inhibited the mycelia growth of other fungi including Botrytis cinerea, Rhizoctonia solani and Alternaria panax, but did not affect the growth of C. destructans. In addition, conidia germination of other fungal species except for C. destructans was inhibited in submerged culture supplemented with radicicol. This medium provides a very efficient tool for isolating C. destructans and also can be used as an enrichment medium for this fungus.

Development of an improved selective media for differentiation of emetic and diarrheal type Bacillus cereus

  • Hong, Yong-Gun;Lee, Jin-Joo;Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.815-818
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    • 2021
  • The objective of this study was to develop a differential medium with improved selectivity for the isolation of Bacillus cereus. Mannitol egg yolk polymyxin medium supplemented with D-galactose allowed the differentiation of diarrheal- and emetic-type B. cereus through pH monitoring. The pH of the medium decreased significantly when incubating the emetic-type B. cereus, whereas the pH change was not significant when incubating the diarrheal-type. The addition of pH indicators, such as methyl red and phenol red, to the medium allowed visual differentiation between diarrheal- and emetic-type B. cereus. A solid agar medium was also developed by optimizing the concentrations of medium components such as monosaccharides, agar, egg yolk enrichment, pH indicators, and antibiotics. This study indicates the possibility of applying selective media for the differentiation of diarrheal- and emetic-type B. cereus.

Recovery and Survival of Listeria monocytogenes in Surface and Sea Water (지표수 및 해수로부터 Listeria monocytogenes의 분리 및 생존성)

  • Yang, Ju;Kim, Toh-Gyong;Kang, Ho-Jo
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.327-333
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    • 2002
  • The study was carried out to examine the distribution and survival rate of Listeria monocytogenes (L monocytogenes) from various source of waters using improved isolation method. In comparision of enrichment media for isolation of L monocytogenes from water, the isolation rate and 50% detection limit of the pathogen were higher in UVM modified Listeria enrichment broth (UVM) than Listeria enrichment broth (LEB). On the other hand, when compared the selective media for isolation of the pathogen from water, the isolation rate was highest in culture at Oxford agar followed by Fraser agar, and LEB agar. In order to improve enrichment method, 100 ml of water samples with 0.1 CFU/ml of L monocytogenes was inoculated into 10 ml of UVM concentrated at 10-fold, and incubated for 24 h at $36^{\circ}C$. Isolated frequency of the pathogens in improved enrichment method completely corresponded with common (filter) method. Of a total mumber of 147 water samples from river, lake and sea, the pathogen was isolated from 1 of 39 (2.6%) river water samples and 1 of 75 (1.3%) sea water samples, but no pathogen was isolated from 33 lake water samples. Serotypes of 2 isolates were identified as type 1. L monocytogenes decreased in number from 7.2-7.4 to 4.2-4.7 log CFU/ml for 1 week poststorage (5 and $20^{\circ}C$), but the pathogens were able to be detected in river and sea water until 8 weeks after storage. However, in tap water, L monocytogenes were decreased to undetectable level after 2 weeks of storage.

Isolation and Characterization of Soil Strains Producing Glutaryl-7-Aminocephalosporanic Acid Acylase

  • Knang, Yong-Ho;Yoo, Ryong-Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.2 no.2
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    • pp.105-108
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    • 1997
  • A search was undertaken to screen microorganisms that produce an enzyme capable of deacylating glutary1-7-amincephalosporanic acid to 7-aminocephalosporanic acid in soil samples. The screening was carried out by preparing enrichment cultures containing glutary-7ACA and cephalosporin C as selective carbon sources. A non-${\beta}$-lactam model compound,, glutary-p-nitroanilide, was synthesized as a substrate suitable for the rapid screening of microorganisms isolated from the enrichment cultures. Two isolates exhibiting acylase activity, designated BY7.4 and BY8.1, were identified as strains of Pseudomonas species. Pseudomonas BY8.1 showed higher acylase activity toward G1-7ACA than Pseudomonas BY7.4. Environmental conditions for the optimal acylase activity of Pseudomonas BY8.1 were shown to be pH9 and 30$^{\circ}C$.

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Ultra-Specific Enrichment of GST-Tagged Protein by GSH-Modified Nanoparticles

  • Lee, Yeon-Ji;Park, Jong-Moon;Huh, Ji-Young;Kim, Min-Sik;Lee, Je-Sun;Palani, Arudra;Lee, Kwang-Yeol;Lee, Sang-Won
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1568-1572
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    • 2010
  • The selective isolation of specific proteins from complex protein mixtures by nanoparticles is reported. Glutathionemodified superparamagnetic nanoparticles were used to purify specific proteins fused with glutathione S-transferase via enzyme-substrate interactions. They demonstrated greatly improved selectivity and efficiency over micron sized capturing beads. The ultra-specific enrichment of target proteins was confirmed by both SDS-PAGE and LC/MS/MS experiments.

On-line SPE-HPLC Method using Alumina Filtering to Selectively Extract Phenolic Compounds from Environmental Water

  • Lee, Sung-Kwang
    • Bulletin of the Korean Chemical Society
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    • v.31 no.12
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    • pp.3755-3759
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    • 2010
  • A on-line SPE (solid phase extraction)-HPLC preconcentration method was developed for the determination of phenolic compounds at trace levels in environmental water sample. XAD-4 and Dowex 1-X8 were used as sorbent in the on-line SPE-HPLC method for the selective enrichment of nine phenolic compounds, which are included in the priority pollutants list of the US EPA. Also alumina prefiltering considerably reduced the amount of interfering peaks due to humic substances that could accumulated due to the preconcentration step and prevent quantification of polar phenolic compounds in environmental water samples. This method was used to determine the phenolic compounds in tap and river water and superiority to the US EPA 625 method in its enrichment factor, pretreatment time, recoveries, and detection limit. The limits of detection were in the range of $0.3-0.9\;{\mu}g/L$ in tap water sample.

Generation of single stranded DNA with selective affinity to bovine spermatozoa

  • Vinod, Sivadasan Pathiyil;Vignesh, Rajamani;Priyanka, Mani;Tirumurugaan, Krishnaswamy Gopalan;Sivaselvam, Salem Nagalingam;Raj, Gopal Dhinakar
    • Animal Bioscience
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    • v.34 no.10
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    • pp.1579-1589
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    • 2021
  • Objective: This study was conducted to generate single stranded DNA oligonucleotides with selective affinity to bovine spermatozoa, assess its binding potential and explore its potential utility in trapping spermatozoa from suspensions. Methods: A combinatorial library of 94 mer long oligonucleotide was used for systematic evolution of ligands by exponential enrichment (SELEX) with bovine spermatozoa. The amplicons from sixth and seventh rounds of SELEX were sequenced, and the reads were clustered employing cluster database at high identity with tolerance (CD-HIT) and FASTAptamer. The enriched nucleotides were predicted for secondary structures by Mfold, motifs by Multiple Em for Motif Elicitation and 5' labelled with biotin/6-FAM to determine the binding potential and binding pattern. Results: We generated 14.1 and 17.7 million reads from sixth and seventh rounds of SELEX respectively to bovine spermatozoa. The CD-HIT clustered 78,098 and 21,196 reads in the top ten clusters and FASTAptamer identified 2,195 and 4,405 unique sequences in the top three clusters from the sixth and seventh rounds, respectively. The identified oligonucleotides formed secondary structures with delta G values between -1.17 to -26.18 kcal/mol indicating varied stability. Confocal imaging with the oligonucleotides from the seventh round revealed different patterns of binding to bovine spermatozoa (fluorescence of the whole head, spot of fluorescence in head and mid- piece and tail). Use of a 5'-biotin tagged oligonucleotide from the sixth round at 100 pmol with 4×106 spermatozoa could trap almost 80% from the suspension. Conclusion: The binding patterns and ability of the identified oligonucleotides confirms successful optimization of the SELEX process and generation of aptamers to bovine spermatozoa. These oligonucleotides provide a quick approach for selective capture of spermatozoa from complex samples. Future SELEX rounds with X- or Y- enriched sperm suspension will be used to generate oligonucleotides that bind to spermatozoa of a specific sex type.

Factors Affecting Foam Separation of Proteins (단백질의 기포분리에 영향을 미치는 요소들에 관한 연구)

  • Lee, Boo-Young;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.220-224
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    • 1987
  • The concentration ranges forming surface excess of bovine serum albumin(BSA) and ovalbumin solutions were determined, and the factors affecting the foam separation of BSA were investigated. The surface tension of BSA solution decreased from 72 to 61 dynelcm, when the concentration changed from $5{\times}10^{-3}$ to $3{\times}10^{-2}%$, and the critical micelle concentration was appeared to be at 0.03% of BSA. At the isoelectric point (pH 4.9) of BSA, the foamate volume was maximum, but enrichment ratio was minimum, resulting in the maximum recovery rate. When the pH deviated from the isoelectric point, the foamate volume decreased and the enrichment ratio increased. The enrichment ratio increased, while the foamate volume decreased drastically as the temperature was elevated above $20^{\circ}C$, resulting in the decrease in recovery rate. As the gas flow rate increased, the enrichment ratio decreased and the foamate volume increased. When $(NH_4)_2SO_4$ was added, the enrichment ratio decreased, but the maximum foamate was obtained at ionic strength 7. The concentration to form the surface excess of ovlbumin, which has lower surface hydrophobicity than BSA, was 200 times higher than that of BSA. This fact indicates the possibility of selective foam separation by hydrophobicity difference of proteins.

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Bacteriological Study of Listeria sp. Isolated from Seawater and Sea Food (해수와 해산물로부터 Listeria 속의 분리와 세균학적 조사)

  • 강치희;이만효;황용일
    • Journal of Life Science
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    • v.13 no.4
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    • pp.390-399
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    • 2003
  • Four species of the genus of Listeria were isolated from seawater and sea food in Kyungnam province, South Korea. These isolated strains were classified into Listeria sp. from different samples by appropriate cultivation conditions and biochemical tests including serological test. In a day enrichment cultivation, the following strains were found out of 100 samples: L. innocua (35%), L. ivanovii (4%), L. monocytogenes (4%), and L. welshimeri (1%). For seven days enrichment culture, L. innocua (38%), L. ivanoii (5%), L. monocytogenes (7%), and L. welshimeri (1%) were isolated. From these results, Listeria species were more efficiently isolated in seven day enrichment broth than in one day enrichment. However, these isolated Listeria species were less grown in the selective medium than in the enrichment medium. Isolation rates of Listeria species showed differency for each sample and Listeria species were more abundantly isolated in shrimps (80%) and crayfishes (80%) than little neck clams (50%), seawater (25%) and mussels (20%). From the results of serological classes for the seven L. monocytogenes, two strains were defined as type I and the other five strains as type IV.

Effect of Dewpoints on Annealing Behavior and Coating Characteristics in IF High Strength Steels Containing Si and Mn (Si, Mn함유 IF 고강도강의 소둔거동 및 도금특성에 미치는 이슬점 온도의 영향)

  • Jeon, Sun-Ho;Shin, Kwang-Soo;Sohn, Ho-Sang;Kim, Dai-Ryong
    • Korean Journal of Metals and Materials
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    • v.46 no.7
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    • pp.427-436
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    • 2008
  • The effect of dewpoints on annealing behavior and coating characteristics such as wettability and galvannealing kinetics was studied by annealing 0.3wt%Si - 0.1~0.4wt% Mn added interstitial-free high strength steels(IF-HSS). The 0.3wt%Si-0.1wt%Mn steel exhibited good wettability with molten zinc and galvannealing kinetics after annealing when the dewpoint of $H_2-N_2$ mixed gas was above $-20^{\circ}C$. It is shown that the wettability and galvannealing kinetics are directly related to the coverage of the external(surface) oxide formed by selective oxidation during annealing. At $N_2-15%H_2$ annealing atmosphere, the increase of dewpoint results in a gradual transition from external to internal selective oxidation. The decrease of external oxidation of alloying elements with a concurrent increase of their subsurface enrichment in the substrate, showing a larger surface area that was free of oxide particles, contributed to the improved wettability and galvannealing kinetics. On the other hand, the corresponding wettability and galvannealing kinetics were deteriorated with the dewpoints below $-20^{\circ}C$. The continuous oxide layer of network and/or film type was formed on the steel surface, leading to the poor wettability and galvannealing kinetics. It causes a high contact angle between annealed surface and molten zinc and plays an interrupting role in interdiffusion of Zn and Fe during galvannealing process.