• Title/Summary/Keyword: Repetitive DNA

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DNA fingerprinting of Brucella abortus isolated from bovine brucellosis outbreaks by repetitive element sequence (rep)-PCR

  • Suh, Dong Kyun
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.199-205
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    • 2005
  • DNA fingerprint patterns of 8 Brucella reference strains and 15 B. abortus field isolates were characterized by repetitive element sequence-based PCR (rep-PCR) using BOX- and ERIC-primers in this study. AMOS PCR differentiated all Brucella field isolates from B. abortus RB51, a vaccine strain by producing a B. abortus-specific 498 bp band. Rep-PCR using BOX-primer produced 13 to 18 bands with sizes of between 230 and 3,300 bp, and discriminated Brucella strains to the species level except B. canis and B. suis. PCR products amplified with ERIC primers were, however, not appropriate for differentiating the Brucella isolates. DNA fingerprint patterns for all B. abortus field isolates were identical among them and were put on one cluster with B. abortus biovar 1 reference strain in the dendrogram, indicating they were highly clonal. These results suggested that rep-PCR using BOX primer might to be a useful tool for calculating genetic relatedness among the Brucella species and for the study of brucellosis epidemiology.

Potential Applications of Recombinant DNA Probes for Relatedness Analysis of Fusarium oxysporum (Fuarium oxysporum의 유연관계 분석을 위한 Recombinant DNA의 Probe로서의 이용 가능성)

  • 김홍기;김영태;유승헌
    • Korean Journal Plant Pathology
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    • v.10 no.1
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    • pp.1-6
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    • 1994
  • Randomly chosen recombinant clones of Fusarium oxysporum were analysed to select useful probes for relatedness analysis of Fusarium oxysporum. Genomic DNA of F. oxysproum f. sp. cubense, digested with HindIII, was ligated to pUC118 and used to transform Escherichia coli strai DH5$\alpha$. Three clones were identified that hybridized to mutiple restriction fragments of some formae speciales of F. oxysporum. These probes detected repetitive sequences in HindIII or EcoRI digested DNAs. Repeated copy clone pFC46, pFC52 and pFC54 showed evident polymorphisms among ten formae speciales of this fungus. Since clone pFC 52 strongly hybridized to multiple EcoRI-digested restriction fragments of f. sp. cubense, it may be useful as a probe for analysis of other genetic characteristics of this forma specialis. The results suggest that our clones might be very useful as probes for relatedness analysis between or within formae speciales of Fusarium oxysporum.

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Cloning and characterization of polyA- RNA transcripts encoded by activated B1-like retrotransposons in mouse erythroleukemia MEL cells exposed to methylation inhibitors

  • Tezias, Sotirios S.;Tsiftsoglou, Asterios S.;Amanatiadou, Elsa P.;Vizirianakis, Ioannis S.
    • BMB Reports
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    • v.45 no.2
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    • pp.126-131
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    • 2012
  • We have previously identified a DNA silent region located downstream of the 3'-end of the ${\beta}^{major}$ globin gene (designated B1-559) that contains a B1 retrotransposon, consensus binding sites for erythroid specific transcription factors and shares the capacity to act as promoter in hematopoietic cells interacting with ${\beta}$-globin gene LCR sequences in vitro. In this study, we have cloned four new non-polyA RNA transcripts being detected upon blockade of murine erythroleukemia (MEL) cell differentiation to erythroid maturation by methylation inhibitors and demonstrated that two of them share high structural homology with sequences of B1 element found within the B1-559 region. Although it is not clear yet whether and how these RNAs interfere with induction of erythroid maturation, these data provide evidence for the first time showing that methylation inhibitors can activate silent repetitive DNA sequences in MEL cells and may have implications in cancer chemotherapy using demethylating drugs as antineoplastic agents.

Detection of DNA Fragment to Differentiate Korean Cattle

  • Yeo, J.S.;Kim, J.W.;Chang, T.K.;Nam, D.H.;Han, J.Y.;Choi, C.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.8
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    • pp.1071-1075
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    • 2002
  • In order to identify and develop the specific DNA marker for the identification of Hanwoo (Korean Cattle) from other breeds, a specific DNA marker of 519 bp was identified and sequenced from polymorphic analysis using RAPD-PCR for 6 cattle breeds. Two different repetitive sequences, $(AAC)_5$ and $(GAAGA)_2$, were selected and designed to use specific probe to develop a DNA marker for Hanwoo specific. When the $(AAC)_5$ probe was applied, the 10 kb specific DNA marker showed in the DNA fingerprinting from 237 of 281 Hanwoo individuals. This novel Hanwoo specific DNA probe is useful to perform the marker-assisted selection for screening Hanwoo purity as an unique genetic source.

Genetic Diversity of Korean Isolates of Pseudomonas tolaasii and WLRO (White Line Reacting Organism) using BOX-, REP-, and ERIC-PCR (BOX-, REP-, ERIC-PCR을 이용한 국내 수집 Pseudomonas tolaasii와 WLRO(White line reacting organism) 균주들의 유전적 다양성)

  • Chee, Hee-Youn;Oh, Se-Jong;Lincoln, S.P.
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.119-123
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    • 1999
  • Genetic diversity of Korean isolates of Pseudomonas tolaasii and WLRO (White line reacting organism) was assessed using BOX-, REP-, and ERIC-PCR analysis. P. tolaasii showed nearly identical band patterns among isolates, whereas considerable DNA polymorphism was found among isolates of WLRO. On the basis of dendogram, WLRO is characterized as a complex group with high degree of genetic differentiation. Genetic relatedness based on repetitive DNA regions was low between P. tolaasii and WLRO isolates.

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Plant genome analysis using flow cytometry

  • Lee Jai-Heon;Kim Kee-Young;Chung Dae-Soo;Chung Won Bok;Kwon Oh-Chang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 1999.05a
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    • pp.162-163
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    • 1999
  • The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.

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Molecular Typing of Vibrio parahaemolyticus by Repetitive Element-PCR (rep-PCR) (Repetitive Element-PCR (rep-PCR)을 이용한 Vibrio parahaemolyticus 의 분자유전학적 아형 분류)

  • Kim, Won Sik;Hong, Seung Bok;Lee, Kyung;Lee, Jung Nam;Shin, Kyeong Seob
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.1
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    • pp.1-6
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    • 2004
  • The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

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Optimization of DNA Extraction from a Single Living Ciliate for Stable and Repetitive PCR Amplification

  • Kim, Se-Joo;Min, Gi-Sik
    • Animal cells and systems
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    • v.13 no.3
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    • pp.351-356
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    • 2009
  • Ciliates are undoubtedly one of the most diverse protozoans that play a significant role in ecology. However, molecular examination, based on comparing the DNA sequences, has been done on a limited number of the species. Because most ciliates are uncultivable and their population sizes are often too small, it is usually difficult to obtain sufficient genomic DNA required for PCR based experiments. In the present study, we evaluated the effectiveness of four commercial DNA extraction procedures that extract high quality genomic DNA from a single ciliate cell. It was discovered that RED Extract-N-$Amp^{TM}$ PCR kit is the best method for removing PCR-inhibiting substances and minimizing DNA loss during purification. This method can also amplify more than 25 reactions of PCR. In addition, this technique was applied to single cells of 19 species belonged to 7 orders under 5 classes that isolated from mixed natural populations. Their small subunit ribosomal DNA (SSU rDNA) was successfully amplified. In summary, we developed a simple technique for the high-yield extraction of purified DNA from a single ciliate cell that may be more useful for rare ciliates, such as tiny and uncultivable marine microbes.

Approximate Periods of Strings based on Distance Sum for DNA Sequence Analysis (DNA 서열분석을 위한 거리합기반 문자열의 근사주기)

  • Jeong, Ju Hui;Kim, Young Ho;Na, Joong Chae;Sim, Jeong Seop
    • KIPS Transactions on Software and Data Engineering
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    • v.2 no.2
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    • pp.119-122
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    • 2013
  • Repetitive strings such as periods have been studied vigorously in so diverse fields as data compression, computer-assisted music analysis, bioinformatics, and etc. In bioinformatics, periods are highly related to repetitive patterns in DNA sequences so called tandem repeats. In some cases, quite similar but not the same patterns are repeated and thus we need approximate string matching algorithms to study tandem repeats in DNA sequences. In this paper, we propose a new definition of approximate periods of strings based on distance sum. Given two strings $p({\mid}p{\mid}=m)$ and $x({\mid}x{\mid}=n)$, we propose an algorithm that computes the minimum approximate period distance based on distance sum. Our algorithm runs in $O(mn^2)$ time for the weighted edit distance, and runs in O(mn) time for the edit distance, and runs in O(n) time for the Hamming distance.