• Title/Summary/Keyword: repetitive sequence.

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Polymorphism and Genetic Relationships Among Magnaporthe grisea Isolates Obtained from Various Hosts by Using Repetitive DNA Sequences (기주가 다른 Magnaporthe grisea 균주간의 Polymorphism과 유전적 유연관계 분석)

  • 김홍기;김영태
    • Korean Journal Plant Pathology
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    • v.12 no.4
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    • pp.389-394
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    • 1996
  • 도열병균, Magnaporthe grisea, 균주간의 유전적 유연관계를 분석하고 그들의 유전에 관한 '기본 정보를 얻고자 DNA polymorphism 분석을 실시하였다. 기주가 다른 도열병 균주들이 공시되었고 cloning에 의해 벼 도열병균 KJ201레이스 균주로부터 생성된 임의 선발 genomic clone들이 공시균주들간의 polymorphism을 밝히기 위해 사용되었던 바 그중 repectitive sequence를 보유한 repeated copy clone 하나가 선발되었다. Clone pMJ6에 의해 밝혀진 repetitive sequence는 Southern hybridization시 벼 분리균주에는 약 30개, 다른 기주 분리균에도 20∼33개의 밴드를 형성하였다. 반면 피 분리균주에는 단지 두 개의 밴드만을 나타내 분리기주가 다른 균주간에 뚜렷한 polymorphism이 존재하였으며 parsimony 분석에서도 역시 아주 먼 cluster를 형성하여 피 분리균은 다른 기주 분리균과 유전적으로 상당히 먼 것으로 추정되었다. 공시균의 genomic DNA를 HindIII로 처리했을 때 pMJ6에 의한 밴드양상은 공시균을 EcoRI으로 처리했을 때의 MGR probe의 밴드 양상과 유사하여 이 repeated copy clone이 도열병균주간의 유전적 유연관계를 분석하는데 MGR 못지않게 유용할 것으로 보인다.

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Genotypic and Phenotypic Diversity of PGPR Fluorescent Pseudomonads Isolated from the Rhizosphere of Sugarcane (Saccharum officinarum L.)

  • Rameshkumar, Neelamegam;Ayyadurai, Niraikulam;Kayalvizhi, Nagarajan;Gunasekaran, Paramsamy
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.13-24
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    • 2012
  • The genetic diversity of plant growth-promoting rhizobacterial (PGPR) fluorescent pseudomonads associated with the sugarcane (Saccharum officinarum L.) rhizosphere was analyzed. Selected isolates were screened for plant growthpromoting properties including production of indole acetic acid, phosphate solubilization, denitrification ability, and production of antifungal metabolites. Furthermore, 16S rDNA sequence analysis was performed to identify and differentiate these isolates. Based on 16S rDNA sequence similarity, the isolates were designated as Pseudomonas plecoglossicida, P. fluorescens, P. libaniensis, and P. aeruginosa. Differentiation of isolates belonging to the same group was achieved through different genomic DNA fingerprinting techniques, including randomly amplified polymorphic DNA (RAPD), amplified ribosomal DNA restriction analysis (ARDRA), repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC), and bacterial repetitive BOX elements (BOX) analyses. The genetic diversity observed among the isolates and rep-PCR-generated fingerprinting patterns revealed that PGPR fluorescent pseudomonads are associated with the rhizosphere of sugarcane and that P. plecoglossicida is a dominant species. The knowledge obtained herein regarding the genetic and functional diversity of fluorescent pseudomonads associated with the sugarcane rhizosphere is useful for understanding their ecological role and potential utilization in sustainable agriculture.

Detection of Mycobacterium leprae by Real-time PCR Targeting Mycobacterium leprae-Specific Repetitive Element Sequence

  • Jin, Hyun-Woo;Wang, Hye-Young;Kim, Jong-Pill;Cho, Sang-Nae;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.16 no.2
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    • pp.127-131
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    • 2010
  • Mycobacterium leprae detection is difficult even with molecular biological techniques due to the low sensitivity of current methodologies. In this report, real-time PCR targeting the M. leprae-specific repetitive element (RLEP) sequence was developed as a new diagnostic tool and evaluated using clinical specimens. For this, M. leprae DNAs were extracted from skin biopsy specimens from 80 patients and analyzed by real-time PCR using TaqMan probe. Then, the detection efficiency of the real-time PCR was compared with that of standard PCR. In brief, the rate of positive detection by the standard PCR and real-time PCR was 32.50% and 66.25%, respectively. The results seemed to clearly show that the TaqMan real-time PCR developed in this study may be a useful tool for sensitive detection of M. leprae from clinical specimens.

Detection of Mycobacterium leprae by Nested PCR Targeting M. leprae-Specific Repetitive Element (RLEP) Sequence

  • Wang, Hye-Young;Kim, Yeun;Bang, Hye-Eun;Kim, Hyun-Chul;Cho, Sang-Nae;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.13 no.1
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    • pp.33-38
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    • 2007
  • The aim of this work was to validate a rapid and an accurate method for detecting Mycobacterium leprae in clinical specimens using nested PCR targeting M. leprae-specific repetitive element (RLEP) sequence. The primers were derived from the RLEP sequence which yield a 272 bp outer product and a 230 bp inner product. The specificity and the sensitivity of the nested PCR were compared with those of single PCR for detecting M. leprae using DNAs isolated from reference strain and various species of Mycobacterium. The results showed that the sensitivity of the nested PCR was about 100 to 1,000 times higher than that of the single PCR and also showed that both the single and the nested PCR were highly specific to M. leprae. Subsequently, the usefulness of the single and nested PCR was evaluated with clinical samples isolated from leprosy patients. The number of positive detections by the single and the nested PCR with a total of 20 specimens from leprosy patients were 9 (45%) and 20 (100%), respectively. The results clearly showed that nested PCR has highest sensitivity in detecting M. leprae from clinical specimens. Therefore, nested primers targeting RLEP sequence developed in this study seems to be useful to detect the presence of M. leprae.

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Development of Restriction Fragment Length Polymorphism(RELP) Markers in Silkworm, Bombyx mori (누에 RFLP(제한단편 다형현상)마커 개발)

  • 고승주;김태산;이영승;황재삼;이상몽
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.96-104
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    • 1997
  • A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

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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.

Repetitive Delivery Scheme for Left and Right Views in Service-Compatible 3D Video Service

  • Yun, Kugjin;Cheong, Won-Sik;Lee, Jinyoung;Kim, Kyuheon;Lee, Gwangsoon;Hur, Namho
    • ETRI Journal
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    • v.36 no.2
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    • pp.264-270
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    • 2014
  • This paper introduces a novel repetitive delivery scheme for the left and right views in service-compatible (SC) 3D video that provides full backward compatibility to a legacy DTV system while retaining HD 3D visual quality without additional bandwidth or a codec over the legacy broadcasting channel. The proposed SC delivery scheme transmits individual view sequences of a 3D video in interlaced form, that is, a left-view sequence of a 3DTV program to be used repeatedly is transmitted first and stored locally, and the right-view sequence of the 3D program is then transmitted. This paper specifically describes the signaling, synchronization, and storage format methods used to validate the proposed SC delivery scheme. The experiment results show that the proposed SC delivery scheme can be effectively applied for an SC 3DTV service without degrading the DTV quality using only legacy DTV platforms.

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.

A Novel Harmonic Identification Algorithm for the Active Power Filters in Non-Ideal Voltage Source Systems

  • Santiprapan, Phonsit;Areerak, Kongpol;Areerak, Kongpan
    • Journal of Power Electronics
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    • v.17 no.6
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    • pp.1637-1649
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    • 2017
  • This paper describes an intensive analysis of a harmonic identification algorithm in non-ideal voltages source systems. The dq-axis Fourier with a positive sequence voltage detector (DQFP) is a novel harmonic identification algorithm for active power filters. A compensating current control system based on repetitive control is presented. A design and stability analysis of the proposed current control are also given. The aim of the paper is to achieve a robustness of the harmonic identification in a distorted and unbalanced voltage source. The proposed ideas are supported by a hardware in the loop technique based on a $eZdsp^{TM}$ F28335 and the Simulink program. The obtained results are presented to demonstrate the performance of the harmonic identification and the control strategy for the active power filter in non-ideal systems.

Genetic Synthesis and Applications of Repetitive Protein Polymers (반복단위 단백질 고분자의 유전공학적 합성 및 응용)

  • Park, Mi-Sung;Choi, Cha-Yong;Won, Jong-In
    • KSBB Journal
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    • v.22 no.4
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    • pp.179-184
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    • 2007
  • This study introduces the characteristics and some applications of repetitive polypeptides, especially to the biomaterial, tissue engineering scaffolds, drug delivery system, and DNA separation systems. Since some fibrous proteins, which consist of repeating peptide monomers, have been reported that their physical properties are changed dramatically by means of temperature alteration or pH shifting. For that reason, fibrous protein-mimetic polypeptides, which are produced by the recombinant technology, can be applied to the diverse biological fields. Repetitive polypeptides can also be used in the bioseparation area such as DNA sequencing, because they make DNA separation possible in free-solution electrophoresis by conjugating DNA fragments to them. Moreover, artificial synthesis of repetitive polypeptides helps to demonstrate the correlations between mechanical properties and structures of natural protein polymer, which have been proven that repetitive domains are affected by the sequence of the repeating domains and the number of repeating subunits. Repetitive polypeptides can be biologically synthesized using some special cloning methods, which are represented here. Recursive directional ligation (RDL) and controlled cloning method (CCM) have been proposed as excellent cloning methods in that we can control the number of repetition in the multimerization of polypeptides and the components of repetitive polypeptides by either method.