• Title/Summary/Keyword: PCR 동정

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Biochemical and Molecular Identification of Antibacterial Lactic Acid Bacteria Isolated from Kimchi (김치에서 항균활성 유산균의 분리 및 동정)

  • Kim, Soo-Young;Kim, Jong-Doo;Son, Ji-Soo;Lee, Si-Kyung;Park, Kab-Joo;Park, Myeong-Soo
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.446-452
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    • 2011
  • Total 480 lactic acid-producing bacteria were isolated from five kinds of kimchi, and their antibacterial activity was tested against Salmonella enterica serovar Typhimurium, Bacillus subtilis, and Pseudomonas aeruginosa using an agar diffusion assay. Among them, 340 isolates showed a halo on MRS agar against one or more indicator strains, which were identified using multiplex PCR, an API 50CHL kit, and a 16S rDNA sequence analysis. As a result, 169 Lactobacillus plantarum, 20 Lactobacillus fermentum, two Lactobacillus paracasei ssp. paracasei, two Lactobacillus sp., and 15 Pediococcus sp. were identified. This may be the first report on the isolation of antibacterial Lactobacillus fermentum from kimchi.

Characteristics and Antimicrobial Resistance Patterns of Staphylococcus aureus Isolated from Horse (국내 말에서 분리된 Staphylococcus aureus의 특성 및 약제 내성 양상)

  • Choi, Seong-Kyoon;Cho, Gil-Jae
    • Journal of Life Science
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    • v.18 no.1
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    • pp.69-74
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    • 2008
  • This study carried out to investigate the antimicrobial resistance and biochemical characteristics of Staphylococcus aureus (S. aureus) isolated from reproductive and respiratory tract in Thoroughbred horse. The specimens were collected from equine vaginal mucosa and upper respiratory tract from March to December 2006 using a culture swab in Korea. S. aureus suspected colonies on blood agar plates were selected and identified as standard biochemical tests and PCR (Applied Biosystems, USA). Antimicrobial resistance test of S. aureus isolates was performed with 30 antimicrobial agents (BBL, USA) by using the agar disk diffusion method. S. aureus isolates were isolated 58 (39.2%) strains of 148 samples: wound 64.7% (11/17), genital discharge 37.0% (37/100) and nasal discharge 32.2% (10/31). Almost isolates showed high resistance to spectinomycin, sulfonamides, erythromycin, tetracyelin, ciprofloxacin and penicillin. These results may provide the basic information to establish strategies for treatment and prevention of reproductive and respiratory disease in Thoroughbred horses in Korea.

Occurrence and Biovar Classification of Bacterial Wilt Caused by Ralstonia solanacearum in Eggplant (Solanum melongena) (가지의 Ralstonia solanacearum에 의한 풋마름병 발생과 생리형의 분화)

  • Lim, Yang-Sook;Lee, Mun-Jung;Cheung, Jong-Do;Rew, Young-Hyun;Kim, Byung-Soo
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.10-14
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    • 2008
  • Batcterial wilt caused by Ralstonia solanacearum is one of important and widespread diseases worldwide as well as in Korea. Bacterial wilt disease caused by R. solanacearum has been reported mainly in solanaceous crops including eggplant (Solanum melongena), tomato (Solanum lycopersicum), potato (S. tuberosum), and pepper (Capsicum annuum). A total of 48 strains of R. solanacearum from eggplant were collected during 2005 and 2006. They were confirmed as R. solanacearum by PCR amplification with primer pair flipcF/flipcR resulting in production of 470-bp DNA fragment. The 15 isolates exhibited pathogenicity on eggplant and tomato, but less virulent on pepper than other species. The biovar of collected isolates, which have been reported of five types worldwide, were classified as biovars 3 and 4 by physiological test. Biovar 4 was the dormant type without pathogenicity on eggplant rootstock, whereas biovar 3 had pathogenicity on eggplant rootstocks that is resistant to R. solanacearum, indicating necessity of breeding new rootstock with resistance to R. solanacearum biovar 3

Genetic Stock Identification of Spotted Flounder, Verasper variegatus from Yeocheun, Korea (범가자미에 대한 유전학적 동정)

  • KIM Kyung Kil;KIM Yoon;NAM Yoon Kwan;KIM Dong Soo
    • Journal of Aquaculture
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    • v.6 no.3
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    • pp.221-233
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    • 1993
  • Cell size, DNA content, chromosome and PCR-based mitochondrial 12S rRNA gene analyses were conducted to obtain basic informations for genetic stock identification of spotted flounder (Verasper variegatus) from Yeocheun, Korea. The mean erythrocytic and nuclear volumes of spotted flounder were $211.10{\mu}m^3$ and $23.03{\mu}m^3$, respectively. The haploid DNA content of this species was 0.79 pg/cell which correspond to $46.5\%$ of carp and to $22.6\%$ of mammals. Spotted flounder had the 2n = 46 acrocentric chromosomes but no heteromorphic sex chromosomes was found. Mitochondrial DNA gene for 12S ribosomal RNA was amplified by polymerase chain reaction (PCR) and the PCR products were subjected to digestion with 15 restriction endonucleases. Restriction enzyme analyses revealed that Ava I, Mae II, Sma I and Xba I had one restriction site in the mitochondrial 12S rRNA gene segment of spotted flounder, while Mae I had two. Segments of 12S rRNA gene from mitochondria in spotted flounder were sequenced and compared with channel catfish and human as controls. The 12S rRNA gene of this species was more similar to that of channel catfish than to human's.

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Antimicrobial Susceptibility and Biochemical Characteristics of Streptococcus suis Isolated from Diseased Pigs in Gyeongbuk Province (경북지방 환돈에서 분리한 Streptococcus suis의 생화학적 성상 및 약제감수성)

  • Choi, Seong-Kyoon;Kim, Seong-Guk;Kim, Young-Hoan;Choi, Jeong-Hye;Jo, Min-Hee;Cho, Gil-Jae
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1549-1555
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    • 2010
  • Streptococcus suis is a worldwide pathogen of a variety of porcine infection and has also been described as a pathogen for humans. We studied biochemical characteristics, antimicrobial susceptibility, and identification of polymerase chain reaction (PCR) of S. suis isolated from diseased pigs in Gyeongbuk province from 2004 to 2009. Sixty-one isolates were identified as S. suis by biochemical characteristics and PCR from 40 farms. The biochemical characteristics of S. suis isolates were production of VP-negative, hippurate, esculin, and arginine decarboxylase-positive, and fermentation of carbohydrate was variable lactose, trehalose, inulin, and raffinose, which was typeable 11 phenotype. In an antimicrobial susceptibility test, the majority of isolates were highly susceptible to amoxicillin/clavulanic acid, ampicillin, cephalothin, cefoperazone and florfenicol, while being highly resistant to streptomycin, kanamycin, amikacin, neomycin, erythromycin, clindamycin, and tetracycline. The isolates were divided into 11 phenotypes of biochemistry. By using PCR, the 16S-rRNA gene DNA fragment was detected at 304 bp from all of isolates. These results may provide the basic information needed to establish strategies for the prevention of S. suis infection in pigs.

The Infection of Heterosporis anguillarum in Cultured Shortfin Eel (Anguilla bicolor pacifica) (양식동남아산 뱀장어(Anguilla bicolor pacifica)의 Heterosporis anguillarum 감염)

  • Kim, Jin-Do;Do, Jeong-Wan;Choi, Hye-Sung;Jo, Hyae-In;Lee, Nam-Sil;Kim, Young-Dae
    • Korean Journal of Environmental Biology
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    • v.32 no.4
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    • pp.382-388
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    • 2014
  • Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

A PCR Denaturing Gradient Gel Electrophoresis (DGGE) Analysis of Intestinal Microbiota in Gastric Cancer Patients Taking Anticancer Agents (PCR-DGGE를 통해 분석한 항암치료에 따른 장내 미생물 변화)

  • Yu, Sun Nyoung;Ahn, Soon Cheol
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1290-1298
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    • 2017
  • Intestinal microbiota is an important factor in the development of immune defense mechanisms in the human body. Treatments with anticancer agents, such as 5-Fluorouracil, Cisplatin, and Oxaliplatin, significantly change the temporal stability and environment of intestinal bacterial flora. The anticancer treatment chemotherapy often depresses the immune system and induces side effects, such as diarrhea. This study investigated the effects anticancer agents have on the intestinal microbial ecosystems of patients with gastric cancer. An exploration of the diversity and temporal stability of the dominant bacteria was undertaken using a DGGE with the 16S rDNA gene. Researchers collected stool samples from patients zero, two and eight weeks after the patients started chemotherapy. After the treatment with anticancer agents, the bacteria strains Sphingomonas paucimobilis, Lactobacillus gasseri, Parabacteroides distasonis and Enterobacter sp. increased. This study focused on the survival of the beneficial microorganisms Bifidobacterium and Lactobacillus in the intestines of cancer patients. The administration of antigastric cancer agents significantly decreased Lactobacillus and Bifidobacterium populations and only moderately affected the main bacterial groups in the patients' intestinal ecosystems. The results showed the versatility of a cultivation independent-PCR DGGE analysis regarding the visual monitoring of ecological diversity and anticancer agent-induced changes in patients' complex intestinal microbial ecosystems.

Polymerase chain reaction for a rapid and specific identification of Streptococcus suis (Streptococcus suis 신속동정을 위한 PCR 기법)

  • Jung, Byeong-yeal;Jung, Suk-chan;Kim, Jong-yeom;Park, Yong-ho;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.771-776
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    • 1998
  • Synthetic oligonucleotide primers of 20 and 21 bases, respectively, were used in the polymerase chain reaction (PCR) to amplify a sequence of the mrp gene, which encodes the muramidase released protein of Streptococcus suis. Amplification was not recorded when 5 other streptococcal species were tested or when 9 different nonstreptococcal species were tested. A DNA fragment of 517bp was amplified from the genomic DNA of S suis. The lower detection limit was 100pg of the genomic DNA. The primers recognized 34 serotypes of S suis reference strains and 9 isolates from pneumonic lung, brain, nasal discharge, tonsil. This results suggest that the amplification of the mrp gene by PCR method is potential for the identification of S suis isolates.

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Identification of Mariner-Like Element(MLE) Gene from Nombyx mori. (누에에서의 Mariner 유사 전이인자유전자의 동정)

  • Lee, Jin-Sung;Hwang, Jae-Sam;Kim, Yong-Sung;Suh, Dong-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.8 no.3
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    • pp.285-293
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    • 1998
  • We have cloned an internal fragment of the putative transoisase gene of MLE in the silkworm, Bombyx mori, using PCR method with degenerative oligonucleotide primers designed to represent regions of amino acids encoding transposase. The resulting PCR clone, designed as BmoMAR, cords a partial ORF(152 a.a.) of MLE in which interrupted by five stop codons, and the sequence of its deduced amino acids showed 37% homology with Mos1, an active mariner, from Drosophila mauritiana. Furthermore, the BmoMAR exhibits nucleotide and amino acid homology with 59% and 37% from Apis mellifera and D. mauritiana 7.9 clone, respectively. This result strongly that a MLE is present in the genome of B. mori.

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Identification of Mycobacteria by Comparative Sequence Apalysis and PCR-Restriction Fragment Length Polymorphism Analysis (염기서열과 PCR-Restriction Fragment Length Polymorphism 분석에 의한 Mycobacteria 동정)

  • Kook, Yoon-Hoh
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.561-571
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    • 1999
  • Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the ${\beta}$ subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.

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