• Title/Summary/Keyword: Molecular identification

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Identification of Nicotiana tabacum Cultivars using Molecular Markers

  • Um, Yu-Rry;Cho, Eun-Jeong;Shin, Ha-Jeong;Kim, Ho-Bang;Seok, Yeong-Seon;Kim, Kwan-Suk;Lee, Yi
    • Journal of the Korean Society of Tobacco Science
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    • v.30 no.2
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    • pp.85-93
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    • 2008
  • This report describes a set of seven informative single-nucleotide polymorphisms (SNPs) and one insertion-deletion (INDEL) distributed over 24 cultivars that can be used for tobacco (Nicotiana tabacum L.) cultivar identification. We analyzed 163,000 genomic DNA sequences downloaded from Tobacco Genome Initiative database and assembled 31,370 contigs and 60,000 singletons. Using relatively long contigs, we designed primer sets for PCR amplification. We amplified 61 loci from 24 cultivars and sequenced the PCR products. We found seven significant SNPs and one INDEL among the sequences and we classified the 24 cultivars into 10 groups. SNP frequency of tobacco, 1/8,380 bp, was very low in comparison with those of other plant species, between 1/46 bp and 1/336 bp. For exact identification of tobacco cultivars, many more SNP markers should be developed. This study is the first attempt to identify tobacco cultivars using SNP markers.

Molecular Identification, Enzyme Assay, and Metabolic Profiling of Trichoderma spp.

  • Bae, Soo-Jung;Park, Young-Hwan;Bae, Hyeun-Jong;Jeon, Junhyun;Bae, Hanhong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1157-1162
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    • 2017
  • The goal of this study was to identify and characterize selected Trichoderma isolates by metabolic profiling and enzyme assay for evaluation of their potential as biocontrol agents against plant pathogens. Trichoderma isolates were obtained from the Rural Development Administration Genebank Information Center (Wanju, Republic of Korea). Eleven Trichoderma isolates were re-identified using ribosomal DNA internal transcribed spacer (ITS) regions. ITS sequence results showed new identification of Trichoderma isolates. In addition, metabolic profiling of the ethyl acetate extracts of the liquid cultures of five Trichoderma isolates that showed the best anti-Phytophthora activities was conducted using gas chromatography-mass spectrometry. Metabolic profiling revealed that Trichoderma isolates shared common metabolites with well-known antifungal activities. Enzyme assays indicated strong cell wall-degrading enzyme activities of Trichoderma isolates. Overall, our results indicated that the selected Trichoderma isolates have great potential for use as biocontrol agents against plant pathogens.

Isolation, Identification, and Characterization of Bacillus strains from the Traditional Korean Soybean-fermented Food, Chungkookjang

  • Joo, Myeong-Hoon;Hur, Sung-Ho;Han, Yong-Soo;Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.202-210
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    • 2007
  • A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

First report of the lichen Ochrolechia akagiensis (Ochrolechiaceae, Ascomycota) in Korea

  • Park, Jung Shin;Oh, Soon-Ok;Woo, Jeong-Jae;Liu, Dong;Park, Sook-Young;Hur, Jae-Seoun
    • The Korean Journal of Mycology
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    • v.47 no.2
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    • pp.95-104
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    • 2019
  • The genus Ochrolechia is a widespread, lichen genus in Korea. Despite being common, little is known about the species diversity and geographical distribution of Ochrolechia. In this study, we detailed the identification procedure of the genus Ochrolechia in a Korean collection and provided the description of each species. Using 104 specimens collected from 2003 to 2017, we identified four species of the genus Ochrolechia via morphological and/or molecular phylogenetic analysis: O. parellula, O. trochophora, O. yasudae and O. akagiensis. Among them, O. akagiensis had not been previously reported in Korea. Moreover, the species identified as O. frigida and O. tartarea in past studies were corrected as O. yasudae and O. parellula, respectively, based on morphological and/or molecular evidence. Phylogenetic analysis using the internal transcribed spacer regions including 5.8S rRNA gene showed that the four species separated clearly, indicating that the morphological identification corresponds to the phylogenetic identification. We provide a taxonomic key for the four species of the genus Ochrolechia.

Non-Invasive Sex Determination of Asiatic Black Bear (Ursus thibetanus) via Sex-Specific Amplification of the Amelogenin Gene

  • Baek-Jun Kim
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.4 no.4
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    • pp.154-158
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    • 2023
  • The Asiatic black bear, Ursus thibetanus, is among the most threatened or endangered species in Asia. For its conservation and management, sex identification of U. thibetanus using non-invasive samples (e.g., hair and/or feces) is potentially valuable. In this study, a non-invasive molecular method for sex identification of U. thibetanus samples collected from various countries was first utilized, and it was based on polymerase chain reaction (PCR) amplification of the amelogenin gene via PCRs. Thirty-three bear DNA samples, extracted not only from blood (n=9) but also from hair (n=18) and feces (n=6), were used. We performed sex-specific PCR amplifications of the amelogenin gene using a primer set, SE47 and SE48. The primer set could successfully amplify a single X-specific band for females and both X- and Y-specific bands for males from all blood (100%) and hair (100%) samples. In addition, the primer set could distinguish the sex of bears in four out of a total of six fecal samples (approximately 67%). This study's findings suggest that this molecular method can be applied to sex identification of Asiatic black bears from various Asian regions using non-invasive samples, such as hair and feces.

Identification of Key beta-1,3-glucan Synthesis Enzymes in Agrobacterium sp. ATCC31750 (Agrobacterium sp. ATCC31750에 대한 beta-l,3-glucan 합성 대사경로의 주요 단백질 검출)

  • Kim Ryo Hwa;Lee Jung Heon
    • KSBB Journal
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    • v.19 no.5
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    • pp.406-409
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    • 2004
  • Matrix Assisted Laser Desorption ionization Time of Flight (MALDI-TOF) was used for enzymes identification related to B -1,3-glucan synthesis. Agrobacterium sp. ATCC31750 was cultivated with two stage Continuous Stirrer Tank Reactor (CSTR) and the cells were harvested and their protein profiles were analysed by two dimensional electrophoresis. The specific enzyme spot was treated with trypsin and ana lysed by MALDI-TOF to get peptide molecular weight. The peptide molecular weights were matched with Agrobacterium tumefacience's Data Base from the matrix science site, then could identify the avaliable key enzymes. In this study, we identified key metabolite of synthesis of beta-1,3-glucan, such as glucose-6-phosphate isomerase, phosphoglucomutase, B-1,3-glucan synthase and glucokinase, and we also identified uracil phosphoribocyl transferase and Ribosome recycling factor also.

In silico target identification of biologically active compounds using an inverse docking simulation

  • Choi, Youngjin
    • CELLMED
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    • v.3 no.2
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    • pp.12.1-12.4
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    • 2013
  • Identification of target protein is an important procedure in the course of drug discovery. Because of complexity, action mechanisms of herbal medicine are rather obscure, unlike small-molecular drugs. Inverse docking simulation is a reverse use of molecular docking involving multiple target searches for known chemical structure. This methodology can be applied in the field of target fishing and toxicity prediction for herbal compounds as well as known drug molecules. The aim of this review is to introduce a series of in silico works for predicting potential drug targets and side-effects based on inverse docking simulations.

First Report of Metacordyceps chlamydosporia (Cordyceps chlamydosporia) Isolated from Soil in Korea

  • Kim, Hyun Seung;Adhikari, Mahesh;Yadav, Dil Raj;Kim, Sang Woo;Um, Yong Hyun;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.48-50
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    • 2016
  • A previously unrecorded species, Metacordyceps chlamydosporia KNU14-22, was isolated from soil in Korea. Identification of the fungal species was based on morphological and molecular characteristics. This species has not been previously reported in Korea and herein we present data with detailed descriptions and figures.

Nano-identification for the Cleavage of Disulfide Bond during the Self-Assembly Processes of Unsymmetric Dialkyl Disulfides on Au(111)

  • Noh, Jae-Geun
    • Bulletin of the Korean Chemical Society
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    • v.26 no.4
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    • pp.553-557
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    • 2005
  • The formation of striped phases of unsymmetric hexyl octadecyl disulfide ($CH_3(CH_2)_5SS(CH_2)_{17}CH_3$, HOD) and 1-hydroxyundecyl octadecyl disulfide ($CH_3(CH_2)_{17}SS(CH_2)_{11}$OH, HUOD) on Au(111) and graphite has been investigated by scanning tunneling microscopy (STM) to understand the self-assembly processes of dialkyl disulfides. STM imaging clearly shows the formation of striped phases having corrugation periodicities that are nearly consistent with the molecular length of alkanethiolate moieties formed after the S-S bond cleavage of dialkyl disulfide on a gold surface. On the other hand, self-assembled monolayers (SAMs) of dialkyl disulfides on a graphite surface displayed long-range, well-ordered monolayers with one striped pattern that shows periodicity as a function of molecular length via nondissociative adsorption. From a nonoscopic viewpoint, we have clearly demonstrated that dialkyl disulfide SAMs on gold form via S-S bond cleavage of disulfide.

Identification and Characterization of Thermoplasma acidophilum 2-Keto-3-Deoxy-D-Gluconate Kinase: A New Class of Sugar Kinases

  • Jung, Jin-Hwa;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.535-539
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    • 2005
  • The thermoacidophilic archaeon Thermoplasma acidophilum has long been known to utilize D-glucose via the non-phosphorylated Entner-Doudoroff (nED) pathway. We now report the identification of a gene encoding 2-keto-3-deoxy-D-gluconate (KDG) kinase. The discovery of this gene implies the presence of a glycolysis pathway, other than the nED pathway. It was found that Ta0122 in the T. acidophilum genome corresponded to KDG kinase. This enzyme shares no similarity with known KDG kinases, and belongs to a novel class of sugar kinases. Of the five sugars tested only KDG was utilized as a substrate.