• Title/Summary/Keyword: DNA fingerprinting

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REP-PCR Genotyping of Four Major Gram-negative Foodborne Bacterial Pathogens (주요 식중독 그람 음성 세균 4속의 REP-PCR genotyping)

  • Jung, Hye-Jin;Seo, Hyeon-A;Kim, Young-Joon;Cho, Joon-Il;Kim, Keun-Sung
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.611-617
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    • 2005
  • Dispersed repetitive DNA elements in genomes of microorganisms differ among and within species. Because distances between repetitive sequences vary depending on bacterial strains, genomic fingerprinting with interspersed repetitive sequence-based probes can be used to distinguish unrelated organisms. Among well-known bacterial repetitive sequences, Repetitive Extragenic Palindromic (REP) sequence has been used to identify environmental bacterial species and strains. We applied REP-PCR to detect and differentiate four major Gram-negative food-borne bacterial pathogens, E. coli, Salmonella, Shigella, and Vibrio. Target DNA fragments of these pathogens were amplified by REP-PCR method. PCR-generated DNA fragments were separated on 1.5% agarose gel. Dendrograms for PCR products of each strain were constructed using photo-documentation system. REP-PCR reactions with primer pairs REP1R-I and REP2-I revealed distinct REP-PCR-derived genomic fingerprinting patterns from E. coli, Salmonella, Shigella, and Vibrio. REP-PCR method provided clear distinctions among different bacterial species containing REP-repetitive elements and can be widely used for typing food-borne Gram-negative strains. Results showed established REP-PCR reaction conditions and generated dendrograms could be used with other supplementary genotyping or phenotyping methods to identify isolates from outbreak and to estimate relative degrees of genetic similarities among isolates from different outbreaks to determine whether they are clonally related.

Conservation of Swertia chirata through direct shoot multiplication from leaf explants

  • Chaudhuri, Rituparna Kundu;Pal, Amita;Jha, Timir Baran
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.213-218
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    • 2008
  • Swertia chirata is an endangered gentian species that prefers to grow at higher altitudes. This ethnomedicinal herb is known primarily for its bitter taste caused by the presence of important phytochemicals that are directly associated with human health benefits. Due to a continuous loss of habitat and inherent problems of seed viability and seed germination, alternative strategies for propagation and conservation are urgently required to prevent the possible extinction of this species. We have formulated a reproducible protocol for the rapid propagation and conservation of this plant using leaves taken from in vitro shoot cultures. Direct induction of more than seven shoot buds per explant was achieved for the first time when the explants were placed on MS medium supplemented with $2.22{\mu}M$ N-6-benzyladenine, $11.6{\mu}M$ kinetin, and $0.5{\mu}M$ ${\alpha}-naphthalene$ acetic acid. Direct organogenesis was noted exclusively from the adaxial surface of the basal segments of leaves. Leaves closer to the apical meristem were more responsive than those farther away from the meristem. Plants raised through direct organogenesis were evaluated for their clonal fidelity by chromosomal analysis and DNA fingerprinting. Complete plants were successfully transferred to the field condition and produced viable seeds. Given the enormous potential of this age-old medicinal plant in terms of potential health-benefitting drugs, this protocol can be used for commercial propagation purposes and to initiate future genetic improvement studies.

Genetic Discrimination of Catharanthus roseus Cultivars by Multivariate Analysis of Fourier Transform Infrared Spectroscopy Data

  • Kim, Suk-Weon;Cho, Soo-Hwa;Chung, Hoe-Il;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.201-205
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    • 2007
  • To determine whether pattern recognition based on metabolite fingerprinting for whole cell extracts of higher plants is applied to discriminate plants genetically, leaf samples of eight cultivars of Catharanthus roseus were subjected to Fourier transform infrared spectroscopy (FT-IR). FT-IR fingerprint region data were analyzed by principal component analysis (PCA). Major peaks as biomarkers were identified as the most significant contributors to distinguish samples by using genetic programming. A hierarchical dendrogram based on the results from PCA separated the eight cultivars into two major groups in the same manner as the dendrograms based on genetic fingerprinting methods such as RAPD and AFLP. A slight difference between the dendrograms was found only in branching pattern within each subgroup. Therefore, we conclude that the hierarchical dendrogram based on PCA of the FT-IR data represents the most probable chemotaxonomical relationship between cultivars, which is in general agreement with the genetic relationship determined by conventional DNA fingerprinting methods.

Use of Terminal Restriction Length Polymorphism (T-RFLP) Analysis to Evaluate Uncultivable Microbial Community Structure of Soil

  • Chauhan, Puneet Singh;Shagol, Charlotte C.;Yim, Woo-Jong;Tipayno, Sherlyn C.;Kim, Chang-Gi;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.127-145
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    • 2011
  • Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

Analysis of Probabilistic Limits of Trait Identity in Inter-Strain Comparison of Genomic Fingerprints of Bacteria (균주간 유전체 지문 비교분석에서 유전형질 일치성의 확률적 한계 분석)

  • Zo, Young-Gun
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.263-267
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    • 2011
  • Genomic fingerprinting methods are useful in determining relatedness among bacterial strains. However, random coincidences in sizes of two DNA fragments in two different fingerprints may occur, resulting in erroneous interpretation of relatedness between two bacterial genomes. In this study, I estimated the probability of occurrence of DNA bands of identical size in fingerprints of two unrelated genomes, so that the significance of fingerprint-based estimation of genome relatedness could be analyzed. The probability could be estimated as outputs of a function formulated with the three parameters: the numbers of observed fragments, all possible sizes of fragments and observed fragments common in a given pair of fingerprints. The parameter most instrumental to significance of relatedness estimation was the number of all possible sizes of fragments. To keep the number of coincidentally-common size of fragments below 10, about 200 fragments should be distinguishable in the fingerprints.

Genetic Identification on Natural Population of Triploid Crucian Carp, Carassius auraus in Korea (자연산 3배체 붕어 (Carassius auratus) 클론 집단에 대한 유전학적 동정)

  • Kim Eung Oh;LEE Jong Yoon;Nam Yoon Kwon;Noh Jae Koo;Lee Sang Yun;Kim Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.589-594
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    • 2002
  • Natural clonal stock of triploid crucian carp, Carassius aurahs was identified and its cytogenetic, molecular genetic and morphological traits were studied. Cytogenetic analysis of the clonal crucian carp revealed that they were natural triploidy, evidenced by 1.5-fold increases of cell size, DNA content, and chromosome number. Multi-locus DNA fingerprinting using $(GATA)_4$ probe showed that they had an identical fingerprint profile, indicating the clonal propagation of the population. External morphology and morphometric characteristics of triploid individuals were much uniform compared to those of diploids. Natural triploid crucian carp was proven to be all-female in this study.

Utilization of DNA Marker-Assisted Selection in Korean Native Animals

  • Yeo, Jong-sou;Kim, Jae-Woo;Chang, Tea-Kyung;Pake, Young-Ae;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.71-78
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    • 2000
  • The recent progress od DNA technologies including DNA fingerprinting (DFP) and random amplified DNA polymorphism (RAPD) analysis make it possible to identify the specific genetic trits of animals and to analyze the genetic diversity and relatedness between or withinspecies or populations. Using those techniquse, some efforts to identify and develop the specific DNA markers based on DNA polymorphism, which are related with economic traits for Korean native animals, Hanwoo(Korean native cattle),Korean native pig and Korean native chicken, have been made in Korea for recent a few years. The developed specific DNA markers successfully characterize the Korean native animals as the unique Korean genetic sources, distinctively from other imported breeds. Some of these DNA markers have been related to some important economic traits for domestic animals, for example, growth rate and marbling for Honwoo, growth rate and back fat thinkness fornative pig, and growth rate, agg weight and agg productivity for native chicken. This means that those markers can be used in important marker-assised selection (MAS) of Korean native domestic animals and further contribute to genetically improve and breed them.

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DNA Profiling of Leuconostoc citreum Strains in Fermented Foods by Repetitive Element Polymerase Chain Reaction

  • Kaur, Jasmine;Sharma, Anshul;Lee, Sulhee;Park, Young-Seo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1778-1782
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    • 2017
  • To identify and discriminate the bacterial species at the subspecific level, rep-PCR is a reliable genomic fingerprinting tool. Fourteen strains of bacteria were isolated from different food sources, identified as Leuconostoc citreum using 16S rRNA gene sequencing, and amplified using rep-primers (REP, ERIC, and $(GTG)_5$). Fingerprinting patterns generated bands in the range of 300-6,000 bp with REP, 150-6,000 bp with ERIC, and 200-1,700 bp with $(GTG)_5$ primers. In UPGMA dendrogram analysis, 14 strains were clustered into three clades (I, II, and III) with all the primers, thus differentiating them at the molecular level. The present study revealed the differentiation of L. citreum strains using rep-PCR.

Fingerprinting of Rice Genomes Using PCR with Arbitrary Primers

  • Park, Kyong-Hee
    • Preventive Nutrition and Food Science
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    • v.3 no.2
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    • pp.198-202
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    • 1998
  • The arbitrary primed polymerase chain reaction (AP-PCR) has been used to detect the genetic alternations in the related species. Simple and reproducible fingerprints of complex genomes can be generated using single arbitrary chosen primers and the PCR. The technique was applied to the Oryza species and characterized the relationship among three cultivars of rice species based on theresult of genomic DNA fingerprints. The results indicated that the polymorphism revealed in rice strains and the differences in the PCR product pattern could be represented for each strainis. There was many variationsin the PCR product pattern between cv. Dongin(japonica type)and cv.Hyangdo (indica type), and our chosen AP-primers can ge as markers for strain identification and verfication.

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