• Title/Summary/Keyword: environmental DNA (eDNA)

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The List of Korean Organisms Registered in the NCBI Nucleotide Database for Environmental DNA Research (환경유전자 연구를 위한 NCBI Nucleotide 데이터베이스에 등록된 국내 생물 목록 현황)

  • Ihn-Sil Kwak;Chang Woo Ji;Won-Seok Kim;Dongsoo Kong
    • Korean Journal of Ecology and Environment
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    • v.55 no.4
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    • pp.352-359
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    • 2022
  • Recently, with the development of genetic technology, interest in environmental DNA (eDNA) to study biodiversity according to molecular biological approaches is increasing. Environmental DNA has many advantages over traditional research methods for biological communities distributed in the environment but highly depends on the established base sequence database. This study conducted a comprehensive analysis of the habitat status and classification at the genus level, which is mainly used in eDNA (12S rRNA, 16S rRNA, 18S rRNA, COI, and CYTB), focusing on Korean registration taxon groups (phytoplankton, zooplankton, macroinvertebrates, and fish). As a result, phytoplankton and zooplankton showed the highest taxa proportion in 18S rRNA, and macroinvertebrates observed the highest ratio in the nucleotide sequence database in COI. In fish, all genes except 18S rRNA showed a high taxon ratio. Based on the Korean registration taxon group, the gene construction of the top 20 genera according to bio density observed that most of the phytoplankton were registered in 18S rRNA, and the most significant number of COI nucleotide sequences were established in macroinvertebrates. In addition, it was confirmed that there is a nucleotide sequence for the top 20 genera in 12S rRNA, 16S rRNA, and CYTB in fish. These results provided comprehensive information on the genes suitable for eDNA research for each taxon group.

Analysis of the characteristics of the environment and fish community in the Gwanggyo Lake Park area using the environmental DNA technique (환경 DNA 기법을 활용한 광교호수공원 일대의 시기 및 수환경 특성별 어류상 분석)

  • Won, Su-Yeon;Kang, Yu-Jin;Song, Young-Keun
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.25 no.5
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    • pp.77-88
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    • 2022
  • This study aims to understand the relationship between the distribution of fish species in the two water ecosystems and the habitat factors according to the survey period targeting Gwanggyo Lake Park in the city. There are studies on the appearance and distribution of species by applying eDNA to freshwater ecosystems. However, in the domestic, streams are the target, and studies on the relationship between species distribution and habitat environment in two water environments are lacking. We conducted to analyze the species list and relationship with habitat factors using eDNA research in May and October at 21 points in Gwanggyo Lake Park, Suwon City, which were connected to lakes and streams. As a result, there was no species difference in the water environment according to the survey period. However, the total number of reads during the spawning season(May) was 3,126,482, which was more than double that after the spawning season(October). Tolerant species appeared in Woncheon Lake with a slow or stagnant flow, but there was no significant correlation between species and habitat factors depending on the survey period. On the other hand, intermediate and sensitive species appeared in the Woncheon stream with high flow. There was a significant correlation between the low temperature during the spawning season and the high dissolved oxygen content after the spawning season(P<0.001, Tem.: 20.7±2.6℃, DO: 8.6±1.7). It is expected that environmental DNA will be used to survey species and suggest monitoring methods according to the survey period.

Evaluation of Genotoxicity in Blood Cells of a Polychaetous Worm (Perinereis aibuhitensis), Using Comet Assay (Comet assay를 이용한 갯지렁이 (Perinereis aibuhitensis)의 혈구세포에 대한 유전독성 평가)

  • Seo Jin Young;Sung Chan Gyoung;Choi Jin Woo;Lee Chang Hoon;Ryul Tae Kwon;Han Gi Myung;Kim Gi Beum
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.333-341
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    • 2005
  • In order to know whether polychaetes could be used as an appropriate organism for the detection of genotoxicity, DNA strand breaks were evaluated in blood cells of a nereidae worm (Perinereis aibuhitensis) exposed to various aquatic chemical pollutants (e.g. Cd, Pb, Pyrene, Benaor[a]pyrene). Hydrogen peroxide increased DNA strand breaks up to the highest concentration (10 $\mu$M). Higher concentration than 0.1 $\mu$M showed a significantly more DNA damage than control. Cadmium and lead also showed higher DNA damage than control, over 1.0 and 1 $\mu$g/L, respectively. In case of pyrene, DNA damage was detected even at 0.001 $\mu$g/L. However, DNA damage decreased due to apoptosis at the highest concentration of pyrene and Pb. This study suggested that the polythaetous blood cells could be used effectively for screening genotoxic contaminants in the environment.

Molecular Cloning and Characterization of a recA-like Gene Induced by DNA Damage from a Fluorescent Pseudomonas sp.

  • Ok Bong Kim;Na Young Kim;Jae Hoon Jeong;Si Wouk Kim;Hye Gwang Jeong;Seong Myeong Yoon;Jong Kun Park;Jung Sup Lee
    • Animal cells and systems
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    • v.3 no.2
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    • pp.229-236
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    • 1999
  • The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

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Physiological Function of a DNA-Binding Protein from Starved Cells in Combating Diverse External Stresses in Escherichia coli (대장균 세포 내 다양한 외부 스트레스에 대한 DPS 단백질의 생리적 기능)

  • Lee, Joo Hyeong;Cheong, Su Jin;Oh, Hun Taek;Kim, Woe Yeon;Jung, Young Jun
    • Journal of Life Science
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    • v.23 no.4
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    • pp.479-486
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    • 2013
  • The DNA-binding protein from starved cells (DPS), originally identified as a DNA binding protein in Escherichia coli, is known to play an important role in DNA protection. The aim of this study was to evaluate the functional roles of DPS in E. coli against various kinds of external stresses by comparing the properties of wild-type E. coli cells and dps knockout mutant E. coli (${\Delta}dps$) cells. Under various stress conditions, we measured the cell growth of the wild-type E. coli and the dps knockout mutant E. coli (${\Delta}dps$) cells using a UV spectrophotometer. The growth rate of the cells was compared to investigate the functional roles of the DPS protein in E. coli. In comparison to the properties of the wild-type E. coli cells, the dps knockout mutant E. coli (${\Delta}dps$) cells showed highly sensitive phenotypes under various stress conditions, such as heat shock, acidic pH, nutrient deficiency, and different concentrations of reactive oxygen species (ROS), suggesting that DPS plays key roles in E. coli in combating diverse external stresses. The DPS DNA-binding protein in E. coli plays crucial roles in bacterial cell growth and in the protection of the cells from environmental stresses by tightly binding and preserving their DNA molecules.

Characterization and functional inferences of a genome-wide DNA methylation profile in the loin (longissimus dorsi) muscle of swine

  • Kim, Woonsu;Park, Hyesun;Seo, Kang-Seok;Seo, Seongwon
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.3-12
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    • 2018
  • Objective: DNA methylation plays a major role in regulating the expression of genes related to traits of economic interest (e.g., weight gain) in livestock animals. This study characterized and investigated the functional inferences of genome-wide DNA methylome in the loin (longissimus dorsi) muscle (LDM) of swine. Methods: A total of 8.99 Gb methylated DNA immunoprecipitation sequence data were obtained from LDM samples of eight Duroc pigs (four pairs of littermates). The reference pig genome was annotated with 78.5% of the raw reads. A total of 33,506 putative methylated regions (PMR) were identified from methylated regions that overlapped at least two samples. Results: Of these, only 3.1% were commonly observed in all eight samples. DNA methylation patterns between two littermates were as diverse as between unrelated individuals (p = 0.47), indicating that maternal genetic effects have little influence on the variation in DNA methylation of porcine LDM. The highest density of PMR was observed on chromosome 10. A major proportion (47.7%) of PMR was present in the repeat regions, followed by introns (21.5%). The highest conservation of PMR was found in CpG islands (12.1%). These results show an important role for DNA methylation in species- and tissue-specific regulation of gene expression. PMR were also significantly related to muscular cell development, cell-cell communication, cellular integrity and transport, and nutrient metabolism. Conclusion: This study indicated the biased distribution and functional role of DNA methylation in gene expression of porcine LDM. DNA methylation was related to cell development, cell-cell communication, cellular integrity and transport, and nutrient metabolism (e.g., insulin signaling pathways). Nutritional and environmental management may have a significant impact on the variation in DNA methylation of porcine LDM.

Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

Evaluation of Genotoxicity of Three Antimalarial Drugs Amodiaquine, Mefloquine and Halofantrine in Rat Liver Cells

  • Farombi E. Olatunde
    • Environmental Mutagens and Carcinogens
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    • v.25 no.3
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    • pp.97-103
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    • 2005
  • The genotoxic effect of antimalarial drugs amodiaquine (AQ), mefloquine (MQ) and halofantrine (HF) was investigated in.at liver cells using the alkaline comet assay. AQ, MQ and HF at concentrations between $0-1000{\mu}mol/L$ significantly increased DNA strand breaks of rat liver cells dose-dependently. The order of induction of strand breaks was AQ>MQ>HF. The rat liver cells exposed to AQ and HF (200 and 400 ${\mu}mol/L$) and treated with (Fpg) the bacterial DNA repair enzyme that recognizes oxidized purine showed greater DNA damage than those not treated with the enzyme, providing evidence that AQ and HF induced oxidation of purines. Such an effect was not observed when MQ was treated with the enzyme. Treatment of cells with catalase, an enzyme inactivating hydrogen peroxide, decreased significantly the extent of DNA damage induced by AQ, and HF but not the one induced by MQ. Similarly quercetin, an antioxidant flavonoid at $50{\mu}mol/L$ attenuated the extent of the formation of DNA strand breaks by both AQ and HE. Quercetin, however, did not modify the effects of MQ. These results indicate the genotoxicity of AQ, MQ and HF in rat liver cells. In addition, the results suggest that reactive oxygen species may be involved in the formation of DNA lesions induced by AQ and HF and that, free radical scavengers may elicit protective effects against genotoxicity of these antimalarial drugs.

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Unusual Mitochondrial DNA Polymorphism of the Blue Mussel (Mytilus edulis) Species Complex on the Southern Coast of Korea

  • Iksoo Kim;Byung-Yoon Min;Myung-Hee Yoon;Myong-Suk Yoo;Doh-Hoon Kim
    • Animal cells and systems
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    • v.3 no.1
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    • pp.79-87
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    • 1999
  • Mitochondrial DNA (mtDNA) from 54 specimens of the blue mussel (Mytilus edulis) species complex sampled from the southern coast of Korea was assayed for polymorphism with a portion of the COIII gene (336 bp). Fifteen haplotypes were found. PAUP, one-step networks, and PHYLIP analyses revealed the presence of two clearly differentiated mitochondrial clades (termed clades B and E), separated by 3.6% of minimum sequence divergence. The distribution pattern of the species appears to be consistent with category II of the phylogeographic pattern sensu (Avise et al., 1987): the presence of two discontinuous and distinct mtDNA genotypes in the same geographic region. This unusual mitochondrial polymorphism was explained by the presence of the Mediterranean species, M. galloprovincialis, possessing mtDNA of both M. galloprovincialis and M. edulis.

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A Thermodynamic Study on the Interaction of Quinolone Antibiotics and DNA

  • Lee, Byung-Hwa;Yeo, Ga-Young;Jang, Kyeung-Joo;Lee, Dong-Jin;Noh, Sang-Gyun;Cho, Tae-Sub
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1031-1034
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    • 2009
  • Fluorescence of quinolones including norfloxacin, ciprofloxacin and S- and R-ofloxacin is quenched upon association with single and double-stranded DNA (ss- and ds-DNA). The ratios of fluorescence intensity in the presence of DNA to its absent were plotted with respect to the DNA concentration to construct the Stern-Volmer plot. The slope of the Stern-Volmer plot become larger as the temperature is lowered, ensuring that the fluorescence quenching is static process, i.e., the fluorescence is quenched by formation of the non-fluorescent complex between quinolone and DNA. In the static quenching mechanism, the quenching constant which is equivalent to the slope of the Stern-Volmer plot, is considered as the equilibrium constant for the association of quinolones and DNA. From the temperature-dependent equilibrium constant, ${\Delta}H^0\;and\;{\Delta}S^0$ was obtained using the van’t Hoff relation. In general, association of the quinolone with ds- as well as ss-DNA is energetically favorable (an exothermic) process while the entropy change was unfavorable. Due to the steric effect of the substituents, the effect of the quinolone ring is smaller on the ss-DNA compared to ds-DNA.