• Title/Summary/Keyword: polymerase chain reaction

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Analysis of Lactobacillus casei and Mutant Strains by Polymerase Chain Reaction (Polymerase Chain Reaction에 의한 Lactobacillus casei 및 돌연변이 균주들의 비교 분석)

  • Nam, Jin-Sik;Lee, Jeong-Jun;Shin, Myeong-Su;Na, Seog-Hwan;Baek, Young-Jin;Yoo, Min
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.577-583
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    • 1994
  • To classify Lactobacillus casei strains on the basis of difference in their chromosomal DNA sequence, we have performed polymerase chain reactions on their chromosomal DNA by using random primers, and followed by analyzing randomly amplified polymorphic DNA fragments. We also developed a mini-preparative method to isolate PCR-grade chromosomal DNA from Lacto- bacillus casei strains within 3 hours. Based on RAPD pattems by polymerase chain reactions with degenerated random primers, 4 Lactobacillus casei strains and 2 mutant strains were successfully discriminated. Results were very sensitive, strain-specific and reproducible. It was also reliable. These results suggest that RAPD may be applied efficiently for the identification of several Lactoba- cillus casei strains.

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Genetic Similarity and Diversity in Crucian Carp(Carassius carassius) Populations by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs

  • Yoon, Jong-Man;Kim, Tae-Sun;Kim, Jong-Yeon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.332-333
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    • 2001
  • Genomic DNA was extracted from the blood of the freshwater crucian carp(Carassius carassius) from Kunsan in Korea, representing genetic similarity by polymerase chain reaction amplification of DNA as twelve of arbitrary primers. The electrophoretic analysis of polymerase chain reaction-random amplified polymorphic DNAs(PCR-RADP) products showed the high levels of similarity between different individuals in crucian carp.

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Comparison of quantitative detection of periodontal pathogens before and after scaling by real-time polymerase chain reaction

  • Kim, Young-Sun;Lee, Jung-Hwa;Lee, Young-Eun
    • Journal of Korean society of Dental Hygiene
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    • v.15 no.6
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    • pp.1063-1071
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    • 2015
  • Objectives: The purpose of the study is to investigate the quantitative detection of periodontal pathogens before and after scaling by real-time polymerase chain reaction. Methods: Participants were voluntarily recruited at D university, and saliva samples were extracted before and after scaling. Multiple real-time polymerase chain reactions were used to analyze characteristics and the amount of nine kinds of periodontal pathogens; Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum, Parvimonas micra, Campylobacter rectus, and Eikenella corrodens. Results: After scaling, most periodontal pathogens except Eikenella corrodens were significantly decreased in all subjects(p<0.05). In addition, the percentage of microorganisms associated with disease, the microorganism risk index of periodontitis and the prevalence of red complex, orange complex, and Aggregatibacter actinomycetemcomitans was also significantly reduced after scaling(p<0.05). Conclusions: Scaling decreased in the amount of major periodontal pathogens and periodontitis prevalence rate.

Use of Molecular Replacement to Determine the Phases of Crystal Structure of Taq DNA Polymerase

  • Kim, Young-Soo;Suh, Se-Won
    • BMB Reports
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    • v.29 no.1
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    • pp.38-44
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    • 1996
  • Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in the polymerase chain reaction method, which is being used for amplifying DNA. Not only does Taq DNA polymerase have high commercial value commercial value for the polymerase chain reaction application, but it is also important in studying DNA replication, because it is apparently an homologue to E. coli DNA polymerase I, which has long been used for DNA replication study (Lawyer et ai., 1993). The crystal structure determination of Taq DNA polymerase was initiated. An X-ray diffraction pattern breaks down a crystal structure into discrete sine waves in a Fourier series. The original shape of a crystal object in terms of electron density may be represented as the sum of those sine waves with varying amplitudes and phases in three dimensions. The molecular replacement method was initially employed to provide phase information for the structure of Taq DNA polymerase. The rotation search using the program MERLOT resulted in a solution peak with 5.4 r.m.s. PC-refinement of the X-PLOR program verified the result and also optimized the orientation angles. Next, the translation search using the X-PLOR program resulted in a unique solution peak with 7.35 r.m.s. In addition, the translation search indicated $P3_121$ to be the true space group out of two possible ones. The phase information from the molecular replacement was useful in the MIR phasing experiment.

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Developing species-specific quantitative real-time polymerase chain reaction primers for detecting Lautropia mirabilis

  • Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.3
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    • pp.140-145
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    • 2021
  • This study aimed to develop Lautropia mirabilis-specific quantitative real-time polymerase chain reaction (qPCR) primers based on the sequence of DNA-directed RNA polymerase subunit beta gene. The PrimerSelect program was used in designing of the qPCR primers, RTLam-F4 and RTLam-R3. The specificity of the qPCR primers were performed by conventional PCR with 37 strains of 37 oral bacterial species, including L. mirabilis. The sensitivity of the primers was determined by qPCR with the serial dilution of purified genomic DNA of L. mirabilis KCOM 3484, ranged from 4 ng to 4 fg. The data showed that the qPCR primers could detect only L. mirabilis strains and as little as 40 fg of genome DNA of L. mirabilis KCOM 3484. These results indicate that this qPCR primer pair (RTLam-F4/RTLam-R3) may be useful for species-specific detection of L. mirabilis in epidemiological studies of oral bacterial infectious diseases such as periodontal disease.

Detection of Extended-Spectrum β-Lactamase Producing Klebsiella pneumoniae by Multiplex Polymerase Chain Reaction (Multiplex Polymerase Chain Reaction을 이용한 Extended-Spectrum β-Lactamase 생성 Klebsiella pneumoniae 균주의 검출)

  • Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.173-178
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    • 2006
  • The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) is the main mechanism of bacterial resistance to third-generation cephalosporins and monobactams, whose prevalence varies depending on the different geographical areas. In the last years it has increased notably to the point of being considered a health problem of great importance. The characterization of the ESBLs producing Klebsiella penumoniae strains present in clinical isolates is time-consuming. I describe here the development of a new system, which consists of a multiplex PCR. I found 51 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disc synergy test showed 47 positive K. pneumoniae, which were K. pneumoniae isolates. All ESBLs producing K. pneumoniae strains were resistant to antibiotic amikacin, gentamicin and ciprofloxacin. By multiplex PCR analysis, $bla_{TEM}$ gene in 17 strains 44 $bla_{SHV}$ genes and $bla_{CTX}$ genes in 33 strains were identified. In this study, the multiplex polymerase chain reaction (PCR) assay was a good method to detect and differentiate ESBLs producing K. penumoniae strains in clinical isolates.

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Production of DNA polymerase from Thermus aquaticus in recombinant Escherichia coli

  • Kim, Sung-Gun;Park, Jong-Tae
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.245-249
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    • 2014
  • Among dozens of DNA polymerases cloned from thermophilic bacteria, Taq DNA polymerase from Thermus aquaticus has been most frequently used in polymerase chain reaction (PCR) that is being applied to gene cloning, DNA sequencing, gene expression analysis, and detection of infectious and genetic diseases. Since native Taq DNA polymerase is expressed at low level in T. aquaticus, recombinant Escherichia coli system was used to produce Taq DNA polymerase in a large amount. Taq DNA polymerase was expressed as a soluble form under the control of tac promoter in E. coli, and purified by heat treatment and ion exchange chromatographies. The purified Taq DNA polymerase was nearly homogeneous and exhibited a similar DNA amplification activity with a commercial Taq DNA polymerase.

Rapid Identification of Lactobacillus plantarium in Kimchi Using Polymerase Chain Reaction

  • Kim, Tae-Woon;Min, Sung-Gi;Choi, Dong-Hun;Jo, Jae-Sun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.881-884
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    • 2000
  • A polymerase chain reaction (PCR) was performed to rapidly identify Lactobacillus plantarum from type strains and kimchi samples. The PCR experiments were carried out using specific oligonucleotide primer sets based on the 16S rRNA gene sequences of L. plantarum. The expected DNA amplificate of 419 bp was obtained when either purified DNA or whole cells of L. plantarum strains reacted with LP primers, yet not with any of the other strains. The PCR product was confirmed by DNA sequencing. Accordingly, since the PCR method used is simple, specific, and rapid, it will be useful for monitoring and evaluation L. plantarum in the mixed microbial population found in kimchi.

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Determination of Enteric Bacteria at Microbiologically Risky Points by Multiplex Polymerase Chain Reaction

  • Mahir Gulec;Bilal Bakir;Recai Ogur;Tekbas, Omer-Faruk
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.327-330
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    • 2002
  • The purpose of this research was to test multiplex polymerase chain reaction in investigating the microbiological quality of the risky surfaces in social living places of a military base where over 15 thousand people live together. In 22 samples of 99, there were no bacteria. Only four of the samples contained Shigella, and one of them contained Salmonella, but 77 of the samples contained thermotolerant coliform organisms. There was no statistically significant difference among the microbiological quality of different sites and different equipment surfaces (p>0.05).