• Title/Summary/Keyword: Environmental clone

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Isolation of N-Iauroyl Tyrosine Antibiotic in E. coli Carrying N-acyl Amino Acid Synthase Gene from Environmental DNA in Korean Soils (한국 토양 환경유래의 N-acyl amino acid synthase 유전자에 의한 대장균 내 항생제 N-lauroyl tyrosine 생산)

  • Yeo, Yun-Soo;Lim, Yoon-Ho;Kim, Jeong-Bong;Yang, Jung-Mo;Lee, Chang-Muk;Kim, Soo-Jin;Park, Min-Seon;Koo, Bon-Sung;Yoon, Sang-Hong
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.262-267
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    • 2007
  • To access the natural product antibiotics produced by uncultured microorganisms, six cosmid libraries of DNA extracted directly from soil samples (environmental DNA, eDNA) were constructed and screened for the production of antibacterial active molecules. Of the approximately 60,000 clones screened, one antibacterial clone (YS92B) was detected. Ethyl acetate extracts of clone YS92B showed antibacterial activity against various pathogenic bacteria (Listeria monocytogenes, Bacillus subtilis, Pseudomonas syringae, Xanthomonas campestris pv. oryzae, Staphylococcus epidemis). Active constituents from cultures of YS92B were isolated and purified using a bioassay-guided fractionation against B. subtilis through a series of procedures (ethyl acetate extraction, Sephadex LH20 column chromatography, High Performance Liquid Chromatography). NMR (Nuclear Magnetic Resonance) spectral analysis of a major antibacterial active YS92B-VII indicated that it is a lauric acid linked to tyrosine. This report describes the characterization of antibacterially active long chain N-acyl derivatives of tyrosine that are produced by eDNA clones hosted in Escherichia coli from Korean soils.

Outbreaks of Imipenem-Resistant Acinetobacter baumannii Producing Carbapenemases in Korea

  • Jeong Seok-Hoon;Bae Il-Kwon;Park Kwang-Ok;An Young-Jun;Sohn Seung-Ghyu;Jang Seon-Ju;Sung Kwang-Hoon;Yang Ki-Suk;Lee Kyung-Won;Young Dong-Eun;Lee Sang-Hee
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.423-431
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    • 2006
  • Among 53 Acinetobacter baumannii isolates collected in 2004, nine imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Korea. Nine carbapenemase-producing isolates were further investigated in order to determine the mechanisms underlying resistance. These isolates were then analyzed via antibiotic susceptibility testing, microbiological tests of carbapenemase activity, pI determination, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. One outbreak involved seven cases of infection by A. baumannii producing OXA-23 ${\beta}-lactamase$, and was found to have been caused by a single ERIC-PCR clone. During the study period, the other outbreak involved two cases of infection by A. baumannii producing IMP-1 ${\beta}-lactamase$. The two clones, one from each of the outbreaks, were characterized via a modified cloverleaf synergy test and an EDTA-disk synergy test. The isoelectric focusing of the crude bacterial extracts detected nitrocefin-positive bands with pI values of 6.65 (OXA-23) and 9.0 (IMP-1). The PCR amplification and characterization of the amplicons via direct sequencing showed that the clonal isolates harbored $bla_{IMP-1}$ or $bla_{oxA-23}$ determinants. The two clones were characterized by a multidrug resistance phenotype that remained unaltered throughout the outbreak. This resistance encompassed penicillins, extended-spectrum cephalosporins, carbapenems, monobactams, and aminoglycosides. These results appear to show that the imipenem resistance observed among nine Korean A. baumannii isolates could be attributed to the spread of an IMP-lor OXA-23-producing clone. Our microbiological test of carbapenemase activity is a simple method for the screening of clinical isolates producing class D carbapenemase and/or class B $metallo-{\beta}-lactamase$, in order both to determine their clinical impact and to prevent further spread.

Molecular Monitoring of Eukaryotic Plankton Diversity at Mulgeum and Eulsukdo in the Lower Reaches of the Nakdong River (낙동강 하류 물금과 을숙도 수환경의 진핵 플랑크톤 종조성에 대한 분자모니터링)

  • Lee, Jee Eun;Lee, Sang-Rae;Youn, Seok-Hyun;Chung, Sang Ok;Lee, Jin Ae;Chung, Ik Kyo
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.3
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    • pp.160-180
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    • 2012
  • We have studied the eukaryotic plankton species diversity to compare the community structure of fresh and brackish waters in the lower reaches of the Nakdong River using metagenomic methods. We constructed 18S rDNA clone libraries of total DNAs extracted from environmental water samples collected at Mulgeum (MG100929, fresh) and Eulsukdo bridge (ES, brackish). Through the steps of colony PCR, PCR-RFLP, sequencing and similarity analysis, we discovered the diverse species composition of eukaryotic plankton. Total 338 clones (170 at MG100929 and 168 at ES) were analyzed, and then we found 74 phylotypes (49 for MG100929 and 25 for ES). From the phylogenetic analysis, we confirmed various eukaryotic plankton of broad range of taxonomic groups, including Stramenopiles, Cryptophyta, Viridiplantae, Alveolata, Rhizaria, Metazoa, and Fungi. We also found several unreported species in Korea and candidates of new taxonomic entities at levels higher than genus. Especially, the cryptic species diversity including unreported phylotypes of Pirsonia (Stramenopiles) and Perkinsea (Alveolata) suggests that the molecular monitoring method can produce new informative biological data in monitoring the changes in the Nakdong River Mouth ecosystem.

Environmental Genomics Related to Environmental Health Biomarker

  • Kim, Hyun-Mi;Kim, Dae-Seon;Chung, Young-Hee
    • Molecular & Cellular Toxicology
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    • v.2 no.2
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    • pp.75-80
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    • 2006
  • Biomarkers identify various stages and interactions on the pathway from exposure to disease. The three categories of biomarkers are those measuring susceptibility, exposure and effect. Susceptibility biomarkers are identifiable genetic variations affecting absorption, metabolism or response to environmental agents. Biomarkers of exposure indicate the amount of a foreign compound that is absorbed into the body. Biological measurements performed on human tissues are vastly expanding the capabilities of classical epidemiology, which has relied primarily on estimates of human exposure derived form chemical levels in the air, water, and other exposure routes. Biomarkers of exposure indicate the amount of a foreign compound that is absorbed into the body. Biological measurements performed on human tissues are vastly expanding the capabilities of classical epidemiology, which has relied primarily on estimates of human exposure derived form chemical levels in the air, water, and other exposure routes. The biomarker response is typical of chemical pollution by specific classes of compound, such as (i) heavy metals (mercury, cadmium, lead, zinc), responsible for the induction of metallothionein synthesis, and (ii) organochlorinated pollutants (PCBs, dioxins, DDT congeners) and polycyclic aromatic hydrocarbons (PAHs), which induce the mixed function oxygenase (MFO) involved in their bio transformations and elimination. Currently genomic researches are developed in human cDNA clone subarrays oriented toward the expression of genes involved in responses to xenobiotic metabolizing enzymes, cell cycle components, oncogenes, tumor suppressor genes, DNA repair genes, estrogen-responsive genes, oxidative stress genes, and genes known to be involved in apoptotic cell death. Several research laboratories in Korea for kicking off these Environmental Genomics were summarized.

Bacterial Community Structure Shifted by Geosmin in Granular Activated Carbon System of Water Treatment Plants

  • Pham, Ngoc Dung;Lee, Eun-Hee;Chae, Seon-Ha;Cho, Yongdeok;Shin, Hyejin;Son, Ahjeong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.99-109
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    • 2016
  • We investigated the relation between the presence of geosmin in water and the bacterial community structure within the granular activated carbon (GAC) system of water treatment plants in South Korea. GAC samples were collected in May and August of 2014 at three water treatment plants (Sungnam, Koyang, and Yeoncho in Korea). Dissolved organic carbon and geosmin were analyzed before and after GAC treatment. Geosmin was found in raw water from Sungnam and Koyang water treatment plants but not in that from Yeoncho water treatment plant. Interestingly, but not surprisingly, the 16S rRNA clone library indicated that the bacterial communities from the Sungnam and Koyang GAC systems were closely related to geosmin-degrading bacteria. Based on the phylogenetic tree and multidimensional scaling plot, bacterial clones from GAC under the influence of geosmin were clustered with Variovorax paradoxus strain DB 9b and Comamonas sp. DB mg. In other words, the presence of geosmin in water might have inevitably contributed to the growth of geosmin degraders within the respective GAC system.

Molecular cloning and expression of glyceraldehyde-3-phosphate dehydrogenase gene under environmental stresses in sweetpotato

  • Kim, Young-Hwa;Song, Young-Sun;Huh, Gyung-Hye
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.95-100
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    • 2008
  • Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a main enzyme in the glycolytic pathway, is involved in cellular energy production and regarded as a housekeeping gene. Previously, cytosolic GAPDH was selected as the most significantly abundant gene in EST library of sweetpotato suspension cells. In this study, a full-length of cDNA clone (IbGAPDH) encoding GAPDH was isolated from suspension-cultured cells of sweetpotato (Ipomoea babatas), and its expression was investigated with a view to understanding the physiological function of GAPDH in relation to environmental stresses. IbGAPDH encoded a 36.9 kDa polypeptide consisting of 337 amino acids. When the deduced amino acid of IbGAPDH was compared with other higher plants, IbGAPDH showed high homology with cytosolic GAPDH. The mRNA level of IbGAPDH significantly increased under environmental stresses, such as $H_2O_2$, MV and cold treatments. Among them, the transcript level of IbGAPDH gene was the highest under cold stress. Further investigation of the transcription level under $10^{\circ}C$ or $15^{\circ}C$ was performed with different tissues of sweetpotato. The transcription of IbGAPDH was increased by cold stress with tissue-specificity, moreover, showed different patterns according to temperature.

IBA Treatment of Poplar Cuttings and Soil Composition Amendment for Improved Adaptability and Survival

  • Cho, Wonwoo;Chandra, Romika;Lee, Wi-young;Kang, Hoduck
    • Journal of Forest and Environmental Science
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    • v.36 no.4
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    • pp.259-266
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    • 2020
  • Poplar trees from the Salicaceae family over the years have been utilized for various reasons which include prevention of deforestation as well as phytoremediation. This study aims to determine the optimal pre-treatment and soil conditions required for propagation of poplar cuttings for increased initial adaptability and survival rate. Five poplar clones (Hanan, 110, 107, DN-34, 52-225) were selected for IBA, soil composition treatments on propagation. IBA pre-treatment of cuttings were utilized 0, 10, and 100 mg l-1 concentrations. Soil compositions were amended with TKS-2+perlite 2:1 (v:v) and sandy clay loam mixed with artificial soil. According to the greenhouse results 10 mg l-1 of IBA showed a significant increase in plant height whereas 100 mg l-1 inhibited plant growth except in clone 110. Soil composition severely affected root growth and hence overall growth of the clones. Sandy clay loam soil had poor to stunted growth compared to TKS-2+perlite.

Eicosapentaenoic Acid (EPA) Biosynthetic Gene Cluster of Shewanella oneidensis MR-1: Cloning, Heterologous Expression, and Effects of Temperature and Glucose on the Production of EPA in Escherichia coli

  • Lee, Su-Jin;Jeong, Young-Su;Kim, Dong-Uk;Seo, Jeong-Woo;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.6
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    • pp.510-515
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    • 2006
  • The putative EPA synthesis gene cluster was mined from the entire genome sequence of Shewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO1600 (acyl transferase, AT), ORFSO1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO1597 (enoyl reductase, ER), ORFSO1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified from S. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genes orfSO1597 and orfSO1604. The DNA fragment was cloned into Escherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in four E. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in the E. coli clone was affected by cultivation temperature, showing maximum yield at $20^{\circ}C$ and no production at $30^{\circ}C$ or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

Analysis of Cone and Seed Characteristics from Different Mating Design Strategies of Pinus densiflora for. multicaulis

  • Lee, Kyungmi;Lee, Hyunseok;Kang, Jun-Won
    • Journal of Forest and Environmental Science
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    • v.37 no.4
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    • pp.309-314
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    • 2021
  • This study aimed to enhance seed productivity and secure genetic resources for Pinus densiflora for. multicaulis. We analyzed the characteristics of cone and seed generated by control pollination between Pinus densiflora (PD) and Pinus densiflora for. multicaulis (PDM). The highest number of cone scales (63.0) was obtained from the self-pollinated (sp) PDM clone B (PDM-sp-B), whereas the lowest number of cone scales (44.7) was obtained from two combinations designated as PDM-A×PD-075 and PDM-A×PD-0111. Both female parents of the hybrids were PDM-A. The highest seed production capacity (80.8) was obtained from the open-pollinated (op) PDM clone B (PDM-op-B). The seed potentials of PDM-B×PD-0111, PDM-op-A, and PDM-sp-B were 67.4, 66.5, and 63.1, respectively. The highest number of fertile scales (41.5) was obtained from PDM-op-B, and the lowest number of fertile scales (28.8) was obtained from PDM-A×PD-075. The total number of aborted ovules and 1st aborted ovules was not statistically significant in the mating design. The cross combination of PDM-B×PD-0111 had the highest number (34.8) of filled seeds and the lowest number of 2nd aborted ovules (5.2) and empty seeds (9). PDM-op-B had the highest number of developed seeds (47.6), although the number of empty seeds was the highest (41.2). Therefore, we conclude that the mating design of PDM-B×PD-0111 is useful for future breeding programs to improve seed yield of PDM. Our results showed that there was a strong correlation between the following two parameter pairs: number of scales and number of fertile scales, and the number of fertility scales and seeds potential (r=0.89 and r=0.84, respectively; both p<0.01).