• Title/Summary/Keyword: environmental DNA

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Development of DNA-Based Assessment Method for Mold in Floor Dust of Dwellings in Korea (바닥 먼지내에서 DNA 기반 곰팡이 분석기법 개발)

  • Lee, Jeong-Sub;Kim, Sung Yeon;Choi, Kil Yong;Ryu, Jungmin;Hwang, Eun Seol;Lee, Juyeong;Kwon, Myunghee;Chung, Hyenmi;Seo, SungChul
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.27 no.4
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    • pp.324-332
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    • 2017
  • Objectives: Much scientific evidence indicate a positive association between moldy environments and respiratory illnesses and/or symptoms. However, few comprehensive assessments of mold have been performed for such settings. Spore counts or microscopic enumeration only may not be sufficient for evaluating fungal exposure. Recently, Mold Specific QPCR technology developed by the US EPA (Environmental Relative Moldiness Index, ERMI) has been widely used worldwide and great performance for assessing fungal exposure has been shown. Methods: We aimed to develop a Korean version of ERMI suitable for the distribution of fungal flora in Korea. Thirty dwellings in the Seoul and Incheon area were selected for sampling, and each was classified as 'Flooded, 'Water-damaged' or 'Non-water-damaged'. Results: Dust on the floor and airborne sampling were collected using an MAS100 and a 'Dustream' collector. Samples were analyzed by quantitative polymerase chain reaction(QPCR) for the 36 molds belonging to ERMI. Student t-test and ANOVA tests were carried out using SAS software. The median ERMI values of flooded, water damaged, and non-water damaged dwellings were 8.24(range: -5.6 to 27.9), 5.47(-25. 4 to 32.7), and -15.30(-24.6 to 14.8), respectively. Significant differences were observed between flooded and non-water damaged dwellings (P=0.001) and between water-damaged and non-water damaged dwellings (P=0.032). Conclusion: Our findings indicate that ERMI values attributed to dust samples in Korea could be applicable for the identification of flooded or water damaged buildings. However, much data is needed for continuously developing the Korean version of ERMI values.

Cloning and Sequencing of the Mitochondrial Cytochrome c Oxidase Subunit II Gene from Rhabditidae Family Nematode (Rhabditidae과 선충의 CO II 유전자 클로닝 및 염기서열 분석)

  • Lee, Sang Mong;Son, Hong Joo;Kim, Keun Ki;Hong, Chang Oh;Park, Hyean Cheal
    • Journal of Environmental Science International
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    • v.28 no.1
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    • pp.75-84
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    • 2019
  • Cytochrome c oxidase subunit II gene(CO II gene) is subunit of cytochrome oxidase, which is complex IV of mitochondria electron transport system. It has been frequently used in molecular phylogenetic studies because the speed of its DNA variation is faster than that of nucleus. It is especially useful in phylogenetic study of molecular biology in insects. In this study, we cloned and sequenced CO II gene of mitochondria DNA from Rhabditidae family nematode. Our results showed that this gene is comprised of 696 base pairs(bp). In the analysis of similarity of this gene with other known genes of 14 species of nematodes in Rhabditida order, we identified that this gene has high similarity with that of Caenorhabditis briggsae(86.0%) and C. elegans(85.6%) in Rhabditidae family. On the meanwhile, it has very low similarity with that of Angiostrongylus cantonensis(31.8%) in Angiostrongylidae family and Metastrongylus salmi(31.6%) in Metastrongylidae family. Based on the results of this study, we suggest that this nematode is closely related with that of Caenorhabditis genus in Rhabditidae family.

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.

Characterization of hrp2 + Gene Related to SNF2 Family in Schizosaccharomyces pombe (Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구)

  • Park, In-Soon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.137-141
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    • 2002
  • The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. In addition, to determine the transcription initiation site of hrp2+ gene, primer extension analysis was performed. This result showed the band of 64 bp. The transcriptional start point was mapped to a position of 47 base pair from the first ATG codon of translational initiation codon. In order to investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of 0.25% Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene.

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Production of Transgenic Birdsfoot Trefoil (Lotus corniculatus L.) Plants by Introduction of E35S Promoter + AtNDPK2 Gene (E35S 프로모터 + AtNDPK2 유전자 도입에 의한 버즈풋 트레포일 (Lotus corniculatus L.) 형질전환체 생산)

  • Kim Ki-Yong;Jang Yo-Soon;Choi Gi-Jun;Sung Byung-Ryeol;Kim Won-Ho;Seo Sung;Lee Byung-Hyun;Kwak Sang-Soo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.26 no.2
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    • pp.83-90
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    • 2006
  • To develop transgenic birdsfoot trefoil (Lotus corniculatus L.) plants tolerant to environmental stress, Arabidopsis NDPK gene (AtNDPK) was introduced into birdsfoot trefoil plants using Agrobacterium-mediated transformation and expressed powerfully under the control of the E35S promoter. The expression vector, pEN-K was used for introduction of AtNDPK gene into birdsfoot trefoil plaits. The transformed calli were selected on kanamycin containing medium and then regenerated. The transformed birdsfoot trefoil plants were cultivated for 4 months on BOi2Y medium. Genomic DNA PCR and Southern blot analysis confirmed the incorporation of AtNDPK into the birdsfoot trefoil genome.

An Assessment of Genotoxicity on Organic Solvent Workers by $^{32}$P-postlabelling Method ($^{32}$P-postlabelling법을 이용한 유기용제 작업장 근로자의 유전독성 평가)

  • 홍대용;김장락;이장호;문중갑;이한우;김동일;박성학;정주화;이홍근
    • Environmental Analysis Health and Toxicology
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    • v.9 no.1_2
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    • pp.37-51
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    • 1994
  • To evaluate the genotoxicities of workers exposed to glue and glue cleaning solution, ambient air monitoring of working place, animal study and human monitoring were carried out. By GC-MS analysis, air samples collected from shoesmaking plant were found to be toluene, xylene, cyclohexane, n-hexane, methyl ethyl ketone, trichloroethylene, butylacetate, isopropyl alcohol. Glue and glue cleaning solution from shoesmaking plant were applicated topically to the CD-1 mice. DNA was isolated from skin 24 hr following the application and analysed for DNA-adducts using the nuclease $P_1$version of $^{32}$P-postlabelling assay. RAL (Relative Adduct Labelling, adducts$10^8$ nucleotides) was significantly increased in a dose-dependent manner in the glue cleaning solution treated mice skin. Peripheral blood DNA-adducts of workers exposed to glue and glue cleaning solution were also analysed by the same method, but there were not significant differences in the peripheral blood DNA-adducts level between exposed and control workers. In addition, glue cleaning solution from shoes factory was evaluated for mutagenicity in the Salmonella plate incorporation assay using strains TA 100 and TA 1535 in the presence and absence of Arochlor 1254-induced rat liver S$_{9}$. There was evident mutagenicity for cleaning solution in TA 100 regardless of $S_9$, but TA 1535 showed positive only in the absence of $S_9$when predicted by Stead model of mutagenicity prediction (p=0.0000). The urine concentrates from workers and controls were also assayed for mutagenicity towards strain TA 100 of Salmonella typhimurium in the presence of $S_9$ using Kado's microsuspension assay, but their mutagenic activities were not found to be significant. These data suggest that shoesmaking workers are exposed to genotoxic compounds and need to be monitored by testing the mutagenicity of human urines. However, $^{32}$P-postlabelling application requires further validation for the routine monitoring of human exposure.osure.

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Phylogenetic Analysis of Culturable Arctic Bacteria

  • Lee, Yoo-Kyung;Kim, Hyo-Won;Cho, Kyeung-Hee;Kang, Sung-Ho;Lee, Hong-Kum;Kim, Yea-Dong
    • Ocean and Polar Research
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    • v.26 no.1
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    • pp.51-58
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    • 2004
  • We isolated and identified culturable Arctic bacteria that had inhabited soils around the Korean Arctic Research Station Dasan located at Ny-Alsund, Svalbard, Norway $(79^{\circ}N,\;12^{\circ}E)$. The collected soils were diluted in distilled water; the diluted soil-water was spread on 3M petri-films at Dasan Station. The petri-films were transported to the laboratory at KORDI, and cultured at $4^{\circ}C$. Colonies grown on the petri-films were subsequently cultured on nutrient agar plates at $4^{\circ}C$ every 7 days. The pure colonies were inoculated into nutrient liquid media, genomic DNA was extracted, and phylogenetic analysis was performed on the basis of 165 rDNA sequences. A total of 227 strains of bacteria were isolated. Among them, 16S rDNA sequences of 185 strains were identical with those of known strains isolated in this study, and 42 strains were finally identified. Phylogenetic analysis using 16S rDNA indicated that the 30 strains belonged to Pseudomonas, 7 strains to Arthrobacter, two strains to Flavobacterium, and the remaining to Achromobacter, Pedobacter, and Psychrobacter. Among the 42 strains, 14 bacteria produced protease: they were 6 strains of Pseudomonax, 4 strains of Arthrobater, an Achromobacter strain, 2 strains of Flavobacterium, and a Pedohacter strain. We expect these Arctic bacteria can be used for screening to develop new industrial enzymes that are active at low temperatures.

Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro (in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • Journal of Environmental Health Sciences
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    • v.24 no.2
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    • pp.126-133
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    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

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Are Cryptic Species Real?

  • Crous, Pedro W.
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.29-29
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    • 2014
  • Since Darwin and Wallace introduced the concept on the evolution of species, scientists have been furiously debating what species are, and how to define them. This basic yet intriguing question has bothered us ever since, as communicating to fellow biologists about fungal species is the very cornerstone of mycology. For the species presently known, this has largely been accomplished via Latin binomials linked to morphology in the absence of DNA barcodes. In recent years mycologists have embraced the ribosomal ITS as official barcode region for Fungi, and this locus is also mainly used in environmental pyrosequencing studies. Furthermore, DNA data can now also be used to describe sterile species in the absence or lack of distinct morphological structures. Recent developments such as the registration of names in MycoBank, and linking the phenotype to the genotype, have significantly changed the face of fungal systematics. By employing the Consolidated Species Concept, incorporating genealogical concordance, ecology and morphology, robust species recognition is now possible. Several international initiatives have since built on these developments, such as the DNA barcoding of holdings of Biological Resource Centres, followed by the Genera of Fungi Project, aiming to recollect, and epitypify all type species of all genera. What these data have revealed, is that most genera are poly- and paraphyletic, and that morphological species normally encompass several genetic entities, which may be cryptic species. Once we provide a stable genetic backbone capturing our existing knowledge of the past 250 years, we will be able to accommodate novelties obtained via environmental sequencing platforms. Being able to communicate these species to other biologists in a clear manner that is DNA-based, will enable scientists to elucidate the importance, role and ecological interactions that these fungi have on our planet.

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DNA-adducts and Exposure to Endocrine Disrupting Chemicals (내분비장애물질에 대한 노출과 DNA-adduct 생성)

  • Yang, Mi-Hee
    • Environmental Analysis Health and Toxicology
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    • v.21 no.4 s.55
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    • pp.375-380
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    • 2006
  • Endocrine disrupting chemicals (EDC) have been emphasized due to their threats in human health. Waste incinerator emission has been emphasized as a source of EDC including polychlorinateddibenzofurans(PCDD/F) and other carcinogenic polycyclic aromatic hydrocarbons (PAHs). Urinary 1-hydroxypyrene (1-OHP) has been used as an exposure biomarker for the PAHs. On the other hand, etheno-DNA adducts, e.g. 1, $N^6-ethenodeoxyadenosine({\varepsilon}dA)$, has been developed as an useful effective or response biomarker for carcinogenesis. Thus, I investigated association between urinary 1-OHP and ${\varepsilon}dA$ levels due to distance from an incinerator which was built more 10 years ago in the middle of a farm in P city. I designated the EDC-high and low exposed group due to distance from the incinerator, i.e. within 2.5km and $5.0{\sim}7.5km$ from the incinerator, respectively. The study subjects were age and sex-matched males and females (mean age, $61.3{\pm}9.6$ yrs; total 40 persons, male, 10; female, 10 for the each group). Urinary 1-OHP and ${\varepsilon}dA$ were analyzed with HPLC-FD and IP-HPLC-FD, respectively. As results, the distance from the incinerator was not associated with urinary 1-OHP nor ${\varepsilon}dA$ levels (p=0.43 and 0.82, respectively). On the other hand, urinary ${\varepsilon}dA$ levels were significantly higher in the hyperlipidemia group (N=10) than normal group (N=30). In conclusion, urinary 1-OHP nor ${\varepsilon}dA$ levels can not be suggested as an incinerator-related exposure nor effective biomarker. However, not only distance from the incinerator bot also systemic approaches including wind and soil contamination are required to assume exposure levels of incinerator-related EDC.