• Title/Summary/Keyword: DNA-based Identification

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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PCR-based Detection of Bovine Papillomavirus DNA from the Cutaneous Papillomas and Surrounding Environments in the Korean Native Cattle, Hanwoo

  • Han, Sang-Hyun;Park, Yong-Sang;Seo, Jong-Pil;Kang, Tae-Young
    • Journal of Veterinary Clinics
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    • v.33 no.6
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    • pp.346-350
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    • 2016
  • Two 1-year old calves of Korean Native cattle (Hanwoo) presented cutaneous papillomas on the face and neck. Type 2 bovine papillomavirus (BPV-2) was identified in the cutaneous papillomas based on BPV-specific PCR and subsequent DNA sequencing analysis results. Using DNA samples extracted from two affected calves and unaffected animals reared in the same stable, BPV-2 was not only detected in the cutaneous papillomas of affected animals based on BPV-specific PCR analysis, but also detected in normal skins, hairs, and their environments based on nested PCR analysis. BPV-2 was also detected in DNA samples isolated from animals and environments of that distinct stable with affected calves. However, no BPV-2 was detected in the drinking water of both stables (infected and unaffected). These findings concluded that BPV-2 was transmitted by direct or indirect contact, not by drinking water. This is the first report to show molecular evidence of BPV-2 infection. Rapid and precise molecular identification can be used to screen BPV-2 in cattle farms to understand the biological roles of BPV in animal diseases.

Characterization of the Complete Mitochondrial Genome of Diphyllobothrium nihonkaiense (Diphyllobothriidae: Cestoda), and Development of Molecular Markers for Differentiating Fish Tapeworms

  • Kim, Kyu-Heon;Jeon, Hyeong-Kyu;Kang, Seokha;Sultana, Tahera;Kim, Gil Jung;Eom, Keeseon S.;Park, Joong-Ki
    • Molecules and Cells
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    • v.23 no.3
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    • pp.379-390
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    • 2007
  • We sequenced and characterized the complete mitochondrial genome of the Japanese fish tapeworm D. nihonkaiense. The genome is a circular-DNA molecule of 13607 bp (one nucleotide shorter than that of D. latum mtDNA) containing 12 protein-coding genes (lacking atp8), 22 tRNA genes and two rRNA genes. Gene order and genome content are identical to those of the other cestodes reported thus far, including its congener D. latum. The only exception is Hymenolepis diminuta in which the positions of trnS2 and trnL1 are switched. We tested a PCR-based molecular assay designed to rapidly and accurately differentiate between D. nihonkaiense and D. latum using species-specific primers based on a comparison of their mtDNA sequences. We found the PCR-based system to be very reliable and specific, and suggest that PCR-based identification methods using mtDNA sequences could contribute to the study of the epidemiology and larval ecology of Diphyllobothrium species.

A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

Identification of a Novel Small Molecule Inhibitor Against SARS Coronavirus Helicase

  • Cho, Jin-Beom;Lee, Jin-Moo;Ahn, Hee-Chul;Jeong, Yong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2007-2010
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    • 2015
  • A new chemical inhibitor against severe acute respiratory syndrome (SARS) coronavirus helicase, 7-ethyl-8-mercapto-3-methyl-3,7-dihydro-1H-purine-2,6-dione, was identified. We investigated the inhibitory effect of the compound by conducting colorimetry-based ATP hydrolysis assay and fluorescence resonance energy transfer-based double-stranded DNA unwinding assay. The compound suppressed both ATP hydrolysis and double-stranded DNA unwinding activities of helicase with IC50 values of 8.66 ± 0.26 μM and 41.6 ± 2.3 μM, respectively. Moreover, we observed that the compound did not show cytotoxicity up to 80 μM concentration. Our results suggest that the compound might serve as a SARS coronavirus inhibitor.

Identification of Irradiated Foods by Using DNA, Immunochemical, and Biological Methods

  • Kim, Kyeung-Eun;Yang, Jae-Seung
    • Preventive Nutrition and Food Science
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    • v.4 no.4
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    • pp.276-282
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    • 1999
  • Ionizing radiation is considered to be an efficient technology to improve food safety and to extend food shelf-life in the food industry, and it has been used in food processing with a number of attributes. Food labeling should be established to enable the consumer to choose food freely, based on label information. A variety of methodologies to determine the physical, chemical, microbiological, and biological changes due to irradiation has been investigated in order to discriminate the irradiated and unirradiated food products for the consumer's free choice in food selection. However, no satisfactory method has been developed so far. In this review, various approaches based on DNA, immunochemical, and biological methods are addressed.

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Korea Barcode of Life Database System (KBOL)

  • Kim, Sung-Min;Kim, Chang-Bae;Min, Gi-Sik;Suh, Young-Bae;Bhak, Jong;Woo, Tae-Ha;Koo, Hye-Young;Choi, Jun-Kil;Shin, Mann-Kyoon;Jung, Jong-Woo;Song, Kyo-Hong;Ree, Han-Il;Hwang, Ui-Wook;Park, Yung-Chul;Eo, Hae-Seok;Kim, Joo-Pil;Yoon, Seong-Myeong;Rho, Hyun-Soo;Kim, Sa-Heung;Lee, Hang;Min, Mi-Sook
    • Animal cells and systems
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    • v.16 no.1
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    • pp.11-19
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    • 2012
  • A major concern regarding the collection and storage of biodiversity information is the inefficiency of conventional taxonomic approaches in dealing with a large number of species. This inefficiency has increased the demand for automated, rapid, and reliable molecular identification systems and large-scale biological databases. DNA-based taxonomic approaches are now arguably a necessity in biodiversity studies. In particular, DNA barcoding using short DNA sequences provides an effective molecular tool for species identification. We constructed a large-scale database system that holds a collection of 5531 barcode sequences from 2429 Korean species. The Korea Barcode of Life database (KBOL, http://koreabarcode.org) is a web-based database system that is used for compiling a high volume of DNA barcode data and identifying unknown biological specimens. With the KBOL system, users can not only link DNA barcodes and biological information but can also undertake conservation activities, including environmental management, monitoring, and detecting significant organisms.

Revisiting Rhytisma lonicericola: Morphological Characterization and Molecular Phylogenetic Analysis

  • Jung, Bok-Nam;Park, Ji-Hyun;Shin, Hyeon-Dong
    • Mycobiology
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    • v.50 no.2
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    • pp.150-154
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    • 2022
  • Rhytisma lonicericola was identified as a tar spot fungus on Lonicera sp. in 1902, and has since been recorded on several species of Lonicera in China, Japan, and Korea. Most of the previous records of R. lonicericola have been based on a list of disease occurrences in the absence of any formal morphological identification or molecular analyses. Using six newly obtained specimens collected in the past 2 years, we confirmed the tar spot fungus found on L. japonica in Korea as R. lonicericola based on morphological examinations and molecular phylogenetic analyses. This fungus was distinguished from R. xylostei, another tar spot fungus on Lonicera, by ascospore size and geographical distributions. We present detailed mycological information and, for the first time, DNA sequence data useful for the identification of R. lonicericola.

Progression of Apoptotic Cells by Pretreatment of Proteinase K

  • Joo, Kyeng-Woong
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.161-165
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    • 2002
  • Apoptosis can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of rat tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. I describe a new modified method for formalin-fixed, paraffin-embedded tissue sections, pretense pretreatment to permeate the tissue sections that involves an TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) is acknowledged as a method of choice in the rapid identification and quantification of the apoptotic cell fraction in paraffin tissue preparations. TUNEL was performed without apoptosis and with apopotosis samples to each of the three concentrations of proteinase K (10, 25, 40 mg/ml) pretreatments. In this study, I show that chemical pretreatments of the tissue sections in proteinase K (25 mg/ml for 15 min at room temperature) considerably enhances the sensitivity of this nick end labelling technique.

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EST-based Identification of Genes Expressed in the Brain of the Olive Flounder Paralichthys olivaceus

  • Lee, Jeong-Ho;Noh, Jae-Koo;Kim, Hyun-Chul;Park, Choul-Ji;Min, Byung-Hwa;Kim, Young-Ok;Kim, Jong-Hyun;Kim, Kyung-Kil;Kim, Woo-Jin;Myeong, Jeong-In
    • Fisheries and Aquatic Sciences
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    • v.12 no.4
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    • pp.286-292
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    • 2009
  • We have constructed a cDNA library using brain samples of olive flounder Paralichthys olivaceus. Here, we described the study on gene identification by screening 356 clones from the brain cDNA library of olive flounder. Here, we screened 356 clones from the library to identify genes. Of these, 176 (49.5%) were identified as orthologs of known genes from olive flounder and other organisms. Among the 176 EST clones, 33 (18.7%) represented 11 unique genes that are identical to expressed sequence tags (ESTs) reported for olive flounder, and 120 (68.2%) represented 102 unique genes known from other organisms. The percentage of unknown genes (50.5%) is higher than in other olive flounder cDNA libraries (Lee et al., 2003, 2006, 2007), reflecting the high complexity of brain tissue. Further studies of expression characterization and developmental behavior related to these genes should provide useful insight into the physiological functions of the brain in olive flounder.