• Title/Summary/Keyword: PCR 동정

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Cloning and Characterization of a Gene for Fibrinolytic Enzyme from Bacillus subtilis BB-1 Isolated from Black Bean Chung-kuk (흑두로 제조한 청국에서 분리된 Bacillus subtillus BB-1으로 부터 혈전용해효소 유전자 크로닝 및 특성규명)

  • Lee Young-Hoon;Lee Sung-Ho;Jeon Ju-Mi;Kim Hong-Chul;Cho Yong-Un;Park Ki-Hoon;Choi Young-Ju;Gal Sang-Wan
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.513-521
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    • 2005
  • A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla I DNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript II SK(-) and showed the fibrinolytic activity to bacterial cells. The Cla I DNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla I DNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and $35^{\circ}C$, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.

Genetic Diversity of Endophytic Fungal Strains Isolated from the Roots of Coastal Plants in Ulleung Island for Restoration of Coastal Ecosystem (해안 생태계 복원을 위한 울릉도에 자생하는 해안식물의 뿌리로부터 분리된 내생진균류의 유전적 다양성 분석)

  • Kim, Miae;You, Young-Hyun;Yoon, Hyeokjun;Kim, Hyun;Seo, Yeonggyo;Khalmuratova, Irina;Shin, Jae-Ho;Lee, In-Jung;Choo, Yeon-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1384-1391
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    • 2012
  • Five coastal plant species, Artemisia fukudo, Aster sphathulifolius, Plantago camtschatica, Sedum oryzifolium, and Setaria viridis, were collected from the coastal region of Ulleung Island (Ulleung-Do, South Korea). Thirty-six endophytic fungi were isolated from the roots of these plants, and all were identified by using PCR with the following specifications: internal transcribed spacer 1 (ITS1), 5.8S rRNA, and ITS2 regions. Phylogenetic analysis indicated that all fungal strains belong to the phylum Ascomycota and comprise four orders (Capnodiales, Eurotiales, Hypocreales, and Pleosporales). Among all the identified species, the Eurotiales species were more abundant than species in the other orders. Nine different genera (Alternaria, Aspergillus, Cladosporium, Exserohilum, Fusarium, Neosartorya, Penicillium, Phoma, and Pyrenochaeta) in the four orders were confirmed. Penicillium and Aspergillus species were the most dominant species among the endophytic fungi isolated from the coastal plants. Shannon's diversity index (H') ranged from 0.684 to 1.609, and the endophytic fungi in S. oryzifolium was more diverse compared to the endophytic fungi in the other plants.

Characteristics of Coagulase-negative Staphylococci Isolates from Dental Clinic Environments in Busan, Korea (부산지역 치과환경에서 분리된 coagulase-negative staphylococci의 특성)

  • Jung, Hye-In;Jung, So Young;Park, Indal;Bae, Il Kwon
    • Journal of Life Science
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    • v.26 no.2
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    • pp.220-225
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    • 2016
  • Coagulase-negative staphylococci (CNS) have recently become the bacteria most frequently found in clinical infections. The aim of this study was to investigate the prevalence, antimicrobial susceptibilities, and molecular characteristics of CNS isolates from dental clinic environments in Busan, Korea. One hundred and fifty-four samples were collected from 10 dental clinics and dental hospitals in Busan from December 2014 to January 2015. Species were identified by matrix-assisted laser desorption/ionization–time-of-flight. Antimicrobial susceptibility was determined by disk diffusion methods. A polymerase chain reaction was performed to detect mecA, mupA gene, and SCCmec types. Of the 154 samples, 10(6.5%) isolates were identified as CNS (5 Staphylococcus epidermidis, 2 Staphylococcus capitis, 2 Staphylococcus, and 1 Staphylococcus haemolyticus). Among the 10 isolates, 6 were resistant to penicillin, 5 were resistant to gentamicin, 3 were resistant to tetracycline, and 2 were resistant to cefoxitin and erythromycin. However, clindamycin, ciprofloxacin, teicoplanin, and trimethoprim-sulfamethoxazole resistant isolates were not present. Genes encoding mecA were detected in 4 (2 S. warneri and 2 S. haemolyticus) isolates, and mupA in 1 (S. epidermidis) isolate. One methicillin-resistant CNS (S. warneri) isolate was determined as being of the SCCmec type I. It is concluded that CNS resistant to various antimicrobial agents was widely distributed in dental clinic environments in Korea.

Phylogenetic Diversity and Community Analysis of Marine Bacteria Associated with Ulva pertusa (구멍갈파래(Ulva pertusa)에 서식하는 해양세균의 계통학적 다양성 및 군집구조 분석)

  • Choi, Ha-Ri;Park, So-Hyun;Kim, Dong-Hwi;Kim, Ji-Young;Heo, Moon-Soo
    • Journal of Life Science
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    • v.26 no.7
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    • pp.819-825
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    • 2016
  • The present study was done to assess the diversity of the bacterial community associated with Ulva pertusa collected from Jeju Island using Restriction Fragment Length Polymorphism (RFLP) marker analysis. For RFLP analysis, a total of 145 bacterial strains associated with Ulva pertusa were screened and cultivated using Marine agar and R2A agar. The PCR amplicons of the 16S rRNA gene from all the isolated strains were digested with HaeIII and RsaI restriction enzymes and then classified into different groups according to their restriction patterns. Strains selected based on the RFLP patterns showed more than 91% 16S rRNA gene sequence similarity when compared with known bacterial species, which include 4 phyla - proteobacteria (alpha-proteobacteria, beta-proteobacteria, gamma-proteobacteria - 63%), firmicutes (11%), actinobacteria (4%), bacteroidetes (22%)–as well as 7 classes (actinobacteria, flavobacteriia, cytophagia, bacilli, α-proteobacteria, γ-proteobacteria, β-proteobacteria), 13 orders, 18 families, and 27 genera. These results confirmed a wide diversity of bacterial communities as contrasted with other regions. The newly isolated 10 strains, which show 16S rRNA sequence similarity of <97% compared to previously identified bacteria, could be noble species. Further experiments, such as morphological, physiological, and biochemical classification, are necessary to confirm the novelty of the newly isolated 10 strains.

Antimicrobial Activity of Maesil (Prunus mume) Extract against Vibrio vulnificus (비브리오 패혈증균에 대한 매실 추출물의 항균활성)

  • Ha, ea-Man;Jeon, Doo-Young;Im, Hyun-Chul;Yoon, Yeon-Hee;Shin, Mi-Yeong;Yoon, Ki-Bok;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.163-169
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    • 2017
  • The purpose of this study was to estimate the antimicrobial activity of Maesil (Japanese apricot, Prunus mume) extract against Vibrio vulnificus. The strains tested in the study were 28 V. vulnificus isolates originated from fish, seawater, mud flat and seawater in fish restaurant. The vvhA gene was detected using real-time PCR and biochemical identification expressed above good identification in 28 isolates of V. vulnificus. All of V. vulnificus used in this study was susceptible to tetracycline and chloramphenicol antibiotics. These two antibiotics were considered to be useful for the treatment of patients. Maesil extracts 2.5% and 5% showed antimicrobial activity against V. cholerae NCCP 13589 and V. parahemolyticus NCCP 11143. V. vulnificus isolate and V. vulnificus NCCP 11135 showed growth inhibition at 1.25%, 2.5% and 5% of Maesil extract, respectively. Compared with V. cholerae and V. parahemolyticus, the antibacterial activity of Maesil extract against V. vulnificus was high. The minimum bactericidal concentration of Maesil extract for V. vulnificus was 1.6%. These results revealed that Maesil extract was found to be very useful for inhibiting the growth of V. vulnificus and can be expected to prevent food poisoning caused by V. vulnificus.

A Study on the Sensory Characteristic of Yogurt and Antimicrobial Activity of Lactobacillus plantarum LHC52 Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum LHC52의 항균활성과 요구르트의 관능성 연구)

  • Lee, Seung-Gyu;Han, Ki-Sung;Jeong, Seok-Geun;Oh, Mi-Hwa;Jang, Ae-Ra;Kim, Dong-Hun;Bae, In-Hyu;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.328-335
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    • 2010
  • The aim of our study was to develop a new starter culture for fermented milk. Polymerase chain reaction screening of 103 acid-producing isolates from Kimchi identified 72 Lactobacillus strains. The ability of the strains to inhibit the growth of the food-borne human pathogens (Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus) was measured, using a conventional paper disk method. Among the 72 strains, strain LHC52 displayed potent antagonistic activity. Use of 16S rDNA sequencing and the API 50CHL system identified the strain as Lactobacillus plantarum and it was designated L. plantarum LHC52. Biochemical analyses revealed especially high antibacterial activity against E. coli. Yogurt produced using L. plantarum LHC52 did not show different microbiological and physicochemical properties compared to conventionally-prepared yogurt, implicating L. plantarum LHC52 as a useful, potently antibacterial starter culture for yogurt preparation.

Assessment of the level and identification of airborne molds by the type of water damage in housing in Korea (국내 주택에서 물 피해 유형에 따른 부유곰팡이 농도 수준 평가 및 동정 분석)

  • Lee, Ju Yeong;Hwang, Eun Seol;Lee, Jeong-Sub;Kwon, Myunghee;Chung, Hyen Mi;Seo, SungChul
    • Journal of odor and indoor environment
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    • v.17 no.4
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    • pp.355-361
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    • 2018
  • Mold grows more easily when humidity is higher in indoor spaces, and as such is found more often on wetted areas in housing such as walls, toilets, kitchens, and poorly managed spaces. However, there have been few studies that have specifically assessed the level of mold in the indoor spaces of water-damaged housing in the Republic of Korea. We investigated the levels of airborne mold according to the characteristics of water damage types and explored the correlation between the distribution of mold genera and the characteristics of households. Samplings were performed from January 2016 to June 2018 in 97 housing units with water leakage or condensation, or a history of flooding, and in 61 general housing units in the metropolitan and Busan area, respectively. Airborne mold was collected on MEA (Malt extract agar) at flow rate of 100 L/min for 1 min. After collection, the samples were incubated at $25^{\circ}C$ for 120 hours. The cultured samples were counted and corrected using a positive hole conversion table. The samples were then analyzed by single colony culture, DNA extraction, gene amplification, and sequencing. By type of housing, concentrations of airborne mold were highest in flooded housing, followed by water-leaked or highly condensed housings, and then general housing. In more than 50% of water-damaged housing, the level of airborne mold exceeded the guideline of Korea's Ministry of Environment ($500CFU/m^3$). Of particular concern was the fact that the I/O ratio of water-damaged housing was greater than 1, which could indicate that mold damage may occur indoors. The distribution patterns of the fungal species were as follows: Penicillium spp., Cladosporium spp. (14%), Aspergillus spp. (13%) and Alternaria spp. (3%), but significant differences of their levels in indoor spaces were not found. Our findings indicate that high levels of mold damage were found in housing with water damage, and Aspergillus flavus and Penicillium brevicompactum were more dominant in housing with high water activity. Comprehensive management of flooded or water-damaged housing is necessary to reduce fungal exposure.

Characterization of Cucumber Mosaic Virus Isolated from Passion Fruit (Passiflora edulis) in Korea (국내 패션프루트(Passiflora edulis)에서 분리한 Cucumber Mosaic Virus의 특성)

  • Ye-Yeong Kim;Tae-Seon Park;Ji-Soo Park;Dong-Joo Min;You-Seop Shin;Jin-Sung Hong
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.60-65
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    • 2024
  • In July 2020, total RNA was extracted from passion fruit (Passiflora edulis) leaves showing virus symptoms such as chlorotic spots and vein banding in Haenam, South Korea. Cucumber mosaic virus (CMV)-HN2 was identified through reverse transcription polymerase chain reaction and sequencing analysis. To confirm the biological characteristics of the CMV infecting passion fruit, 10 indicator plants were inoculated with CMV-HN2, and the results showed a typical CMV symptoms. Phylogenetic analysis based on the amino acid of the coat protein (CP) of CMVs revealed that the CMV passion fruit isolates belonged to subgroup I, among which CMV-HN2 belonged to subgroup IA. Additionally, CMVs isolated from passion fruit in Korea have amino acid sequence variation between the subgroup. Among them, CMV-HN2 had four to eight amino acid differences in CP from other CMV isolates from passion fruit. These results confirm the presence of genetic diversity in the CPs of passion fruit CMV isolates.

Distribution Survey of Babesia and Assessment of Tick-borne Diseases in Jeju, Republic of Korea (Babesia의 제주 지역 내 분포 조사)

  • Jiro KIM;YoungMin YUN
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.3
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    • pp.189-197
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    • 2024
  • From March to November 2021, a study conducted in Jeju used the dry ice trap method to collect 17,855 ticks across six regions, examining their distribution and potential as disease vectors. The ticks were identified and categorized by species and growth stage. In addition, the study focused on Babesia, a disease transmitted by hard ticks. Of 17,641 ticks from which DNA had been extracted, 581 pools underwent polymerase chain reaction testing. Of these, 43 pools tested positive for Babesia, with the highest positivity found in Western Seogwipo (23 pools, 53.5%), followed by eastern Seogwipo and central Jeju. The peak times for positive results were April and July. This study highlights an increased risk of tick bites linked to the rising number of abandoned pets in Jeju, necessitating ongoing environmental management and monitoring. These findings provide fundamental data for formulating strategies to prevent and manage tick-borne diseases in Jeju. This involves reducing the disease incidence through targeted preventive measures and detailed epidemiological research. These results underscore the necessity for continued vigilance and proactive intervention to address the health challenges ticks pose in the region.

A Study Bioremediation of Tidal Flat by Microorganism in Pilot Scale Test (환경정화 미생물에 의한 갯벌의 생물학적 정화에 대한 파일럿 규모의 연구)

  • Choi, Hye Jin;Han, Young Sun;Park, Doo Hyun;Oh, Bo Young;Hur, Myung Je;Jo, Nam-Gyu;Kim, Young Hee;Kim, Jong-Guk
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1110-1117
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    • 2014
  • Tidal flats are continuously contaminated by human activities. This study assessed the bioremediation efficiency of tidal flat soil using microcosm reactors and microorganisms originating from the tidal area. We screened 135 bacterial strains that produce extracellular enzymes from the tidal area located in the North port of Incheon bay. Two bacterial strains (Pseudoalteromonas sp. and IC35 Halothiobacillus neapolitanus IC_S22) were selected and used in the microcosm reactors, which were specially designed to functionally mimic the ecological conditions of the tidal flats. Pseudoalteromonas sp. IC35 was selected based on its relatively high activity of the enzymes amylase, cellulose, lipase, and protease. Halothiobacillus neapolitanus IC_S22 was selected for oxidation of sulfur. The M1 and M2 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M2 was first inoculated with Pseudoalteromonas sp. IC35 before the seawater feeding. The initial COD in both the M1 and M2 microcosm reactors was 320 mg/l. The final COD was 21 mg/l (M1) and 7 mg/l (M2). The M3 and M4 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M4 was first inoculated with H. neapolitanus IC_S22. The initial sulfate concentration in both the M3 and M4 microcosm reactors was 660 mg/l, and the maximum sulfate concentration was 1,360 mg/l (M3) and 1,600 mg/l (M4).