• Title/Summary/Keyword: highly repetitive sequence

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Cloning and Characterization of Highly Repetitive Sequences in the Genome of Allium sativum L. (마늘(Allium sativum L.) 게놈의 고반복서열의 분이와 특성 조사)

  • 이동희
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.49-55
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    • 1996
  • We have studied the DNA of Allium sativum L. with respect to highly repetitive sequences. Fast reassociated DNA fragments expected to be highly repetitive sequences based on $C_{o}t$ curve were isolated and characterized. Their copy numbers were approximately $10^{5}~10^{7}$ per haploid genome. Nucleotide sequences analysis of six candidates reveals that their G/C content were low, 25-40% and typical patterns of repeating sequences exist. Repeat sequences were used as probes to access restriction fragment length polymorphism (RFLP) of genomic DNAs of four local clones, Tanyang, Mungyong, So san, and Uisong. The hybridization pattern were very similar among these four local clones.clones.

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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.

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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Molecular characterization of a repetitive element of Xanthomonas oryzae pv. oryzae

  • Yun, Choong-Hyo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.1-19
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    • 1995
  • The plasmid pJEL 101 contains a highly repetitive element from the genome of Xanthomonas oryae pv. oryzae that has properties of an insertional element. The insertional nature of the element, hereto referred to as IS203, was confirmed by molecular analyses of the element and three related elements that were isolated from X. oryzae. The related sequences were isolated on the basis of transposition to the transposon-trapping vector pL3SAC and hybridization with pJEL101. The trapped elements (IS203a, IS203b, and IS203c) were each composed of 1,055 base pairs with 25 base terminal inverted repeats. The elements caused a three base pair target site duplication at the site of insertion in the sacRB gene. The sequence of pJEL 101 has 96% base pair identity with IS203a and 99% identity with IS203a and IS203c but lacks three nucleotides of the consensus left terminal repeat. IS203b has the same DNA sequences as IS203c but is inserted ito the sacRB gene in the opposite orientation. The longest open reading frame of IS203a could code for a protein of 318 amino acids and molecular weight of 37, 151. A search of the Genbank database revealed that IS203 has 51% identity with 909 nucleotides of IS4551 from Escherichia coli. The predicted protein of ORF1 has 40% and 30% amino acid identity to the ORF1 of Tn4551 and the transposase of IS30, respectively.

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Cloning, Sequencing and Characterization of Mitochondrial Control Region of the Domestic Silkwom, Bombyx mori

  • Lee, Jin-Sung;Kim, Ki-Hwan;Hoe, Hyang-Sook;Park, Jae-Heung;Kang, Seok-Woo;Lee, Sang-Han;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.87-89
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    • 2001
  • The nucleotide sequence of the domestic silkworm (Bombyx mori) mitochondrial (mt) control region and its flanking genes was determined from PCR clones. The control region of the silkworm mt genome was located between the small ribosomal RNA gene and transfer RN $A^{Met}$. This 499 bp control region hale 95.4% A+T content. Extensive comparative analysis studies performed with similar control region of other insect genomes could not reveal a highly conserved region containing conserved motifs of animal mito-chondrial genome. The remarkable feature that found in this control region was the presence of tandem motifs containing nine repetitive sequences. The potential usefulness of this motif sequences for Bombyx species or their taxonomically related species is enhanced by its unique localization in the maternally inheritance mitochondrial molecule.e.

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Comparison of Different PCR-Based Genotyping Techniques for MRSA Discrimination Among Methicillin-Resistant Staphylococcus aureus Isolates

  • Kim, Keun-Sung;Seo, Hyun-Ah;Oh, Chang-Yong;Kim, Hong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.788-797
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    • 2001
  • The usefulness of three PCR methods were evaluated for the epidemiological typing of Staphylococcus aureus: an enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), repetitive extragenic palindromic element PCR (REP-PCR), and 16S-23S intergenic spacer PCR (ITS-PCR). The analysis was performed using a collection of S. aureus strains comprised of 6 reference and 79 isolates from patients with various diseases. Among the 85 S. aureus strains tested, 6 references and 6 isolates were found to be susceptible to methicillin, whereas the remaining 73 isolates were resistant to it. PCR methods are of special concern, as conventional phenotypic methods are unable to clearly distinguish among methicillin-resistant S. aureus (MRSA) strains. The ability of the techniques to detect different unrelated types was found to be as follows: ERIC-PCR, 19 types; REP-PCR, 36 types; and ITS-PCR, 14 types. On the basis of combining the ERIC, REP, and ITS fingerprints, the 85 S. aureus strains were grouped into 56 genetic types (designated G1 to G56). The diversities for the 85 S. aureus strains, calculated according to Simpson\`s index, were 0.88 for an ERIC-PCR, 0.93 for a REP-PCR, and 0.48 for an ITS-PCR, and the diversity increased up to 0.97 when an ERIC-PCR and REP-PCR were combined. The above discrimination indices imply that the genetic heterogeneity of S. aureus strains is high. Accordingly, this study demonstrates that DNA sequences from highly conserved repeats of a genome, particularly a combination of ERIC sequences and REP elements, are a convenient and accurate tool for the subspecies-specific discrimination and epidemiologic tracking of S. aureus.

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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.

Evolutionary course of CsRn1 long-terminal-repeat retrotransposon and its heterogeneous integrations into the genome of the liver fluke, Clonorchis sinensis

  • Bae, Young-An;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • v.41 no.4
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    • pp.209-219
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    • 2003
  • The evolutionary course of the CsRn1 long-terminal-repeat (LTR) retrotransposon was predicted by conducting a phylogenetic analysis with its paralog LTR sequences. Based on the clustering patterns in the phylogenetic tree, multiple CsRn1 copies could be grouped into four subsets, which were shown to have different integration times. Their differential sequence divergences and heterogeneous integration patterns strongly suggested that these subsets appeared sequentially in the genome of C. sinensis. Members of recently expanding subset showed the lowest level of divergence in their L TR and reverse transcriptase gene sequences. They were also shown to be highly polymorphic among individual genomes of the trematode. The CsRn1 element exhibited a preference for repetitive, agenic chromosomal regions in terms of selecting integration targets. Our results suggested that CsRn1 might induce a considerable degree of intergenomic variation and, thereby, have influenced the evolution of the C. sinensis genome.

Rapid Detection Methods for Food-Borne Pathogens in Dairy Products by Polymerase Chain Reaction (PCR 방법을 이용한 우유 및 유제품에서 발생하는 식중독 균의 신속 검출법)

  • Kwak, Hyelim;Han, Seonkyeong;Kim, Eiseul;Hong, Yeun;Kim, Haeyeong
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.171-177
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    • 2013
  • The dairy industry has consistently grown via the expansion of dairy-based food categories. Dairy product consumption is stable since the nutrient composition in dairy products is ideal for human health. However, dairy products are highly susceptible to food-borne pathogens. Controlling the safety of dairy products is thus important when considering the nutrient-rich matrix of this food category. Currently, immunoassays or molecular biology techniques have been used to evaluate the safety of dairy products in Korea. These methods are based on the detection of proteins and thus have low reproducibility and sensitivity. Recent techniques to detect food-borne pathogens have focused on genetic analyses. Rapid detection methods for food-borne pathogens in milk and dairy products using polymerase chain reaction (PCR) techniques, such as conventional PCR, real-time PCR, repetitive sequence-based (rep)-PCR, PCR-denaturing gradient gel electrophoresis (DGGE), and digital PCR, are reviewed in this article. The aim of this review was to contribute knowledge of the relationship between microflora and the quality of dairy products. This study will also assist in the immediate monitoring of food-borne pathogens in milk and dairy products when an outbreak related to this food category occurs.

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