• Title/Summary/Keyword: PCR 동정

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Growth Inhibition of Listeria monocytogenes by Weissella spp. from Kimchi Through Real-time PCR (실시간 정량 PCR을 통한 김치 유래 Weissella spp.에 의한 Listeria monocytogenes 생육 억제)

  • Lee, Young-Duck;Kim, Dae-Yong;Park, Jong-Hyun
    • Journal of Food Hygiene and Safety
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    • v.30 no.1
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    • pp.103-108
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    • 2015
  • Weissella spp. from traditional Korean foods of Kimchi were isolated and characterized against food-borne pathogenic Listeria monocytogens. The isolates were identified as W. cibaria 0D17 and W. confusa 0D23 from Kimchi by the biochemical characteristics and 16S DNA sequencing. The culture solutions of the isolates adjusted to pH 7.0 showed L. monocytogens inhibition. To analyze the quantitative detection of L. monocytogenes, real-time PCR was performed according to the SYBR Green I method. The isolates grew well and L. monocytogens did not grow during the co-culture with those strains at $37^{\circ}C$. Therefore, W. cibaria 0D17 and W. confusa 0D23 might be the candidates as the functional lactic acid bacteria for improving food safety.

Characterization of Cucumber mosaic virus Isolated from Trifolium repens in Korea (국내 토끼풀에서 분리한 Cucumber mosaic virus의 특성)

  • Park, Tae Seon;Choi, Gug Seoun;Hong, Jin Sung
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.55-58
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    • 2016
  • A Cucumber mosaic virus (named CMV-Tr1) isolated from the white clover (Trifolium repens) showing mosaic and malformation that found in a pepper field. Cucumber mosaic virus was identified through confirmation with PT-PCR, PCR-restriction fragment length polymorphism, and sequence analysis of coat protein (CP) gene. CMV-Tr1 mosaic symptom on the upper leaves of five tobacco species including Nicotiana benthamiana, Cucumis sativus, Physalis angulata, and Solanum lycopersicon. In Chenopodium quinoa and Vigna unguiculata the isolate showed local lesions in inoculated leaves. CMV-Tr1 compared with CMV-As in the sequence identity of CP gene. CMV-Tr1 showed 98.9% and 99.5% homologies at nucleotide and amino acid levels, respectively. Phylogenetic analysis of the CP gene indicated that CMV-Tr1 belongs to the CMV subgroup IB base on the CP. To our knowledge, this is the first report of CMV in T. repens in Korea.

Phylogenetic Relationships Using ITS2 Sequence and RAPD-PCR Data from Four Species of Korean Pseudo-nitzschia (Bacillariophyceae) (ITS2 부위의 염기서열 및 RAPC-PCR에 의한 Pseudo-nitzschia 4종의 유연관계)

  • Cho, Eun-Seob;Lee, Young-Sik
    • Journal of Life Science
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    • v.14 no.1
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    • pp.32-37
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    • 2004
  • A portion of ribosomal internal transcribed spacer (ITS) 2 was sequenced from the samples of Pseudo nitzschia (P. deticatissima, P. multiseries, P. pungens and P. subfraudulenta) to investigate the genetic characteristics by measuring tile magnitude of genetic diversity and the degree of similarity coefficient using random amplified polymorphic DNAs (RAPD)-PCR patterns. The phylogenetic trees inferred from the genetic distance analyses showed the placement of P. delicatissima formed a quite long distance from p. P. multiseries, P. pungens, and even P. subfraudulenta. The phylogenetic tree from RAPD patterns showed that P. multiseries and P. pungens had dissimilarity coefficient of 0.31, while P. delicatissima and three species of Pseudo-nitzschia had that of 0.81. It is likely thought that the genetic position of P. delicatissima formed far from P. multiseries, P. punges, and P. subfraudulenta. These results imply that ITS2 region is expected to support a useful molecular characters for recognizing at the species level and for even discriminating P. multiseries from P. pungens. RAPD method also will be used to differentiate the species of Pseudo-nitzschia in a short time.

Genetic Diversity of Ralstonia solanacearum Strains Isolated from Pepper and Tomato Plants in Korea (우리나라에 분포하는 고추와 토마토 풋마름병균(Ralstonia solanacearum) 계통들의 유전적 다양성)

  • Seo, Sang-Tae;Park, Jong-Han;Han, Kyoung-Suk;Cheong, Seung-Ryong;Lee, Seung-Don
    • Research in Plant Disease
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    • v.13 no.1
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    • pp.24-29
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    • 2007
  • A total of 35 strains of Ralstonia solanacearum isolated from wilted pepper and tomato plants in Korea were analyzed for their genetic diversity by bacteriological, pathological and molecular biological approaches. All the strains were identified as R. solanacearum biovar 4 on the basis of physiological and biochemical tests, and species-specific PCR primers. Pathogenicity of the strains was confirmed by inoculating on 4-week-old pepper and tomato seedlings. Using cluster analysis based on repetitive sequence-based polymerase chain reaction (rep-PCR) genomic fingerprints, R. solanacearum strains isolated from pepper and tomato in Korea divided into 6 groups showing a high degree of genetic diversity at 55% similarity level. The genetic diversify of strains was not significantly correlated with their geographic origins and host plants.

Evaluation of the preservation state of human skeletal remains using real-time PCR (출토 인골 DNA의 real-time PCR 정량에 의한 보존상태 평가 연구 - 부여 오수리 출토 인골을 중심으로 -)

  • Kwon, Eun-Sil;Cho, Eun-Min;Kim, Sue-Hoon;Kang, Soyeong
    • 보존과학연구
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    • s.32
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    • pp.171-183
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    • 2011
  • In this study molecular genetic analysis was carried out on 4 human skeletal remains from Osuri, Buyeo. We showed that real-time PCR is the method of the choice to assess the initial number of genuine ancient DNA molecules. Human mitochondrial DNA quantification was accomplished by the real-time PCR for the cytochrome b gene of the mitochondria. Histological results proved to be a good potentiality for biochemical analysis using biomolecule. The level of specimen's preservation state was proved that level of quantitative result was BO-04, BO-01, BO-03, BO-02. Continually, we showed that biochemical and biomolecule results for the level of preservation state were similar. This study will be useful to important material for predicting biochemistry and biology analysis of the ancient bone.

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Identification and Characteristics of a Purple, Non-Sulfur Bacterium Rhodobacter sp. EGH-24 from Korea Coast

  • 김기한;차미선;이나은;이정은;이상준;박재림
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11b
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    • pp.233-235
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    • 2003
  • 한국 서해안과 남해안 47개소의 해수와 mud 시료로부터 광합성세균으로 유추되는 13개의 균주를 분리하였고, agar-shake tube method와 RAPD-PCR을 이용하여 4개 서로 다른 균주를 순수분리 하였다. 4개의 균주 중 폐수분해능이 가장 뛰어난 균을 선정하여, 형태학적, 배양적, 생화학적 특성 및 16S rRNA sequencing에 의한 동정결과 purple, sulfur bacteria 쪽에 가까웠으나 형태학적, 배양학적, 생화학적 특성이 purple, non-sulfur bacteria의 Rhodobacter 속에 가장 근접하여, Rhodobacter sp. EGH-24로 명명하였다.

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양식넙치의 연쇄구균병 원인균인 Streptococcus iniae의 실험감염

  • 이주석;지보영;이남실;하은미;심두생
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.322-323
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    • 2003
  • 연쇄구균병은 담수어 및 해산어에 패혈증을 일으켜 양식산업에 막대한 피해를 주는 질병이다. 해산양식어류인 방어(Kusuda et al., 1976; Minami, et al., 1979), 넙치(Nagatsugawa, 1983; Heo et al., 2001), 터봇(Domenech et al., 1996), gilthead sea bream 및 European sea bass (Zlotkin et al., 1998a) 등의 연쇄구균병에 관한 연구가 활발히 진행되고 있고, 특히 최근에는 PCR에 의한 신속진단 및 동정에 관한 연구가 이루어지고 있다(Aoki et al., 2000; Lee et al., 2001; Nguyen et al., 2002). (중략)

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Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR (Polymerase Chain Reaction을 활용한 국내 동물과 사람환자에서 분리한 Staphylococcus aureus 분리주의 분자역학적 특성분석)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.24-37
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    • 2005
  • This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.

Molecular Diagnosis of Grapholita molesta and Grapholita dimorpha and Their Different Occurrence in Peach and Plum (복숭아순나방과 복숭아순나방붙이의 분자동정법 개발 및 복숭아와 자두에서의 발생차이)

  • Ahn, Seung-Joon;Choi, Kyung-Hee;Kang, Taek Jun;Kim, Hyung Hwan;Kim, Dong-Hwan;Cho, Myoung Rae;Yang, Chang Yeol
    • Korean journal of applied entomology
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    • v.52 no.4
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    • pp.365-370
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    • 2013
  • The plume fruit moth, Grapholita dimorpha Komai, a fruit tree pest occurring in the northeast Asia, was firstly reported to infest apple in Korea in 2009, but its direct damage to other fruit trees has been poorly studied. In this study, we investigated shoots and fruits of both peach and plum trees and compared their damage rates by G. dimorpha to those by G. molesta, a congeneric species. In order to discriminate the two moth species, we developed a molecular diagnosis method using species-specific primer sets on different PCR conditions and distinguished the two species collected from the damaged shoots or fruits. The shoots and fruits of peach were infested mostly by G. molesta. However, in plums, the shoots were damaged by G. molesta and the fruits mostly by G. dimorpha. In addition, these two species showed a clear difference in host preference in fruit damage, where 92.5% of the Grapholita moths collected in peach fruits were identified as G. molesta, but 97.0% of the moths in plum fruits were G. dimorpha. The difference of the damage between the two fruit trees may give important information for monitoring of the two moth species in these orchards.

Screening and Identification of Soy Curd-Producing Lactic Acid Bacteria (두유 커드를 생산하는 김치 유래 젖산균의 동정)

  • Kim, Ro-Ui;Ahn, Soon-Cheol;Yu, Sun-Nyoung;Kim, Kwang-Youn;Seong, Jong-Hwan;Lee, Young-Guen;Kim, Han-Soo;Kim, Dong-Seob
    • Journal of Life Science
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    • v.21 no.2
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    • pp.235-241
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    • 2011
  • The purpose of this study was to isolate soy curd forming bacterial strains. Soy curd forming bacteria were isolated from Kimchi, a traditional Korean vegetable food that is fermented using lactic acid bacteria. Among 196 bacterial strains, ten isolates (strain No. 2-2-2, 2-15-2, 2-18-1, 2-19-2, 3-4-1, 3-4-2, 3-8-1, 3-8-3, 3-17-1, 4-39-5) formed firm soy curd. The isolated bacterial strains were identified by molecular biological and biochemical analyses. The genomic DNAs extracted from the isolated bacterial strains were used as a template for PCR amplification of 16S rDNA region. By comparing the results of the 16s rDNA sequences with GenBank data, the isolated strains were identified as Leuconostoc mesenteroides group and Lactobacillus sakei group. The phylogenetic position of soy curd forming strains and their related taxa were investigated using neighbor-joining method. L. mesenteroides group was further identified as L. mesenteroides subsp. dextranicum based on biochemical properties. L. sakei group was named Lactobacillus sp., because it showed a variety of biochemical properties.