• Title/Summary/Keyword: 환경DNA

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Horizontal Gene Transfer among Bacteria by Transformation in Soil and Aquatic Environments (토양 및 수계환경에서 Transformation에 의한 세균들간의 수평적 유전물질 전이)

  • 이건형
    • Korean Journal of Environmental Biology
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    • v.18 no.2
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    • pp.205-213
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    • 2000
  • Laboratory studies have revealed that naturally transformable bacteria develop competence under in situ conditions. Thus, the occurrence of competent bacteria in the environment can be considered as a certainty The persistence of free DNA in natural habitats is influenced by nucleolytic degradation and protection from degradation by adsorption to minerals. Although DNA seeded into natural environment was hydrolysed at substantial rates, but was still detectable at low levels after even several weeks. Compared to the number of laboratory based studies, only a few data have been published dealing with transformation of bacteria in the field. Recently, the potential transfer of recombinant DNA (rDNA) from deliberately or accidentally released bacteria to indigenous microbes has raised biosafety issues, since the persistence of rDNA becomes independent of the survival of its original host and leads to unpredictable, long-term ecological effects. The aim of the present review is to summarise recent literature on horizontal gene transfer (HGT) by transformation among bacteria in both soil and aquatic habitat and special emphasis is placed on recent reports which have addressed HGT among bacteria in the field. [Transformation, Horizontal gene transfer (HGT), recombinant DNA (rDNA), Genetically modified microorganisms (GMMs), Biosafety]

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Enteric Virus Detection from Environmental Sample by Oligonucleotide DNA Chip (올리고뉴클레오티드 DNA Chip을 이용한 환경시료에서의 장관계바이러스 검출)

  • 김정미;윤성욱;지영미;윤재득;정용석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.186-191
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    • 2002
  • The usefulness of oligonucleotide DNA chip was evaluated for detection and primary level identification of major waterborne viruses in environmental samples. The enteric waterborne viruses included enterovirus, adenovirus, and rotavirus. Total intracellular RNA of 10 BGM cell plates showing virus-specific cytopathic effects was extracted at the third day after inoculation. The intracellular RNA was then subjected to either enterovirus-specific RT-PCR followed by sequencing analysis, or the DNA chip. Seven out of 10 positive samples in cell culture were positive but the other three sample were turned out to be negative by both RT-PCR and DNA chip analyses. Nucleotide sequencing results and the DNA chip hybridization results of the RT-PCR product were in complete agreement in the identification of the 7 positive samples as enteroviruses. Using the DNA chip, it took only 3∼4 hr to complete detection and primary level identification of target viruses and additional procedures such as gel electrophoresis or nucleotide sequencing were not necessary. We believe that the DNA chip system can be employed as a highly effective and new detection methodology for environmental viruses.

Effect of 3-Aminobenzamide on DNA Repair Synthesis and Chromosome Aberrations Induced by Mutagens in Synchronized Mammalian Cells (동시화된 포유동물세포에서 돌연변이원에 의해 유발된 DNA 회복합성 및 염색체이상에 미치는 3-Aminobenzamide의 영향)

  • 신은주;강인영;엄경일
    • Environmental Mutagens and Carcinogens
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    • v.11 no.2
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    • pp.107-117
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    • 1991
  • The effect of 3-aminobenzamide (3AB), an inhibitor of poly (ADP-ribose) polymerase, on ethyl methanesulfonate (EMS)-or bleomycin (BLM)-induced DNA repair synthesis and chromosome aberrations was examined during the cell cycle of Chinese hamster ovary (CHO)-K$_1$ cells. The synchronized cells were obtained by using thymidine double block method and mitotic selection method. Three assays were employed in this study: unscheduled DNA synthesis, alkaline elution and chromosome aberrations. 3AB alone did not induce DNA repair and chromosome aberrations in all phases. The post-treatment with 3AB inhibited DNA repair synthesis induced by EMS or BLM in G$_2$ phase, whereas 3AB did not affect chromosome aberrations induced by EMS or BLM in all phases. These results suggest that 3AB aggravates the cell cycle disturbance which occur after DNA damage, and leads to an accumulation of cells at G$_2$ phase, and inhibits DNA repair synthesis, while the effect 3AB on chromosome aberrations may need reevaluated.

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Bacterial Virus DNA Damage Caused by Fumonisin B1 (Fumonisin B1에 의한 세균바이러스 DNA손상)

  • 이길수;조성국
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.34-38
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    • 1999
  • Fumonisin B1 is a secondary metabolite of Fusarium moniliforme, a contaminant of corn and corn product. Fumonisin B1 has been shown to be responsible for major toxicological effects of the fungus in rats, horses, and pigs. Fumonisin B1 induced λ DNA fragmentation, which was increased with incubation time, reducing agent NADPH and metal ion (Cu2+). The DNA damage was inhibited by dimethyl sulfoxide (DMSO) or mannitol as radical scavenger for free radicals. DNA fragmentation, induced by fumonisin B1 in the presence of 1 mM NADPH and 0.1 mM CuCl2, was inhibited by 100 mM DMSO. By the in vitro reaction of fumonisin B1 with supercoiled plasmid pBR322 DNA, plasmid DNA was relaxed, eventually linearized in the agarose gel electrphoresis. From rifampicin sensitive E. coli CSH138 in bacterial mutagenesis system, the rifampicin resistant E. coli mutants were obtained by fumonisin B1. These results suggest that fumonisin B1 may be a possible environmental mutagen in bacterial mutagen assay system.

Effect of the Internal Shape of eDNA Filter Case made by the PBF method Metal 3D Printer on Water Sampling Performance (PBF 방식 금속 3D프린터로 제작된 환경DNA 필터 케이스의 내부 형상이 포집 성능에 미치는 영향)

  • Lee, Seung-Min;Park, Se-Hyun;Kwak, Ihn-Sil;Kim, Hyoung-Ho;Kwak, Tae-Soo
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.20 no.8
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    • pp.74-79
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    • 2021
  • This study focuses on designing a filter case using a water pump for application in eDNA filtering systems. Filter cases, channel type and net type were designed based on the flow field and made using a 3D printer for metal. Flow analysis was conducted for each filter case, and the results were consistent with the pressure experiment results. Furthermore, the water sampling performance test showed that the channel-type filter case exhibited a high flow rate and low pressure through the filter. The eDNA extraction experiment showed that the channel type exhibited improved capture ability compared to the net type.

Feasibility of Environmental DNA Metabarcoding for Invasive Species Detection According to Taxa (분류군별 외래생물 탐지를 위한 환경 DNA 메타바코딩 활용 가능성)

  • Yujin Kang;Jeongeun Jeon;Seungwoo Han;Suyeon Won;Youngkeun Song
    • Journal of Environmental Impact Assessment
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    • v.32 no.2
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    • pp.94-111
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    • 2023
  • In order to establish an effective management strategy for invasive species early detection and regular monitoring are required to assess their introduction or dispersal. Environmental DNA (eDNA) is actively applied to evaluate the fauna including the presence of invasive species as it has high detection sensitivity and can detect multiple species simultaneously. In Korea, the applicability evaluation of metabarcoding is being conducted mainly on fish, and research on other taxa is insufficient. Therefore, this study identified the feasibility of detecting invasive species in Korea using eDNA metabarcoding. In addition, to confirm the possibility of detection by taxa, the detection of target species was evaluated using four universal primers (MiFish, MiMammal, Mibird, Amp16S) designed for fish, mammals, birds, and amphibians. As a result, target species (Trachemys scripta, 3 sites; Cervus nippon, 3 sites; Micropterus salmoides, 7 sites; Rana catesbeiana, 4 sites) were detected in 17 of the total 55 sites. Even in the selection of dense sampling sites within the study area, there was a difference in the detection result by reflecting the ecological characteristics of the target species. A comparison of community structures (species richness, abundance and diversity) based on the presence of invasive species focused on M.salmoides and T.scripta, showed higher diversity at the point where invasive species were detected. Also, 1 to 4 more species were detected and abundance was also up to 1.7 times higher. The results of invasive species detection through metabarcoding and the comparison of community structures indicate that the accumulation of large amounts of monitoring data through eDNA can be efficiently utilized for multidimensional ecosystem evaluation. In addition, it suggested that eDNA can be used as major data for evaluation and prediction, such as tracking biological changes caused by artificial and natural factors and environmental impact assessment.

Detection of Mutated DNA Fragment by the Heteroduplex Analysis at the Temperature Gradient Gel (온도 기울기(temperature gradient) 젤에서 Heteroduplex Analysis 기법을 이용한 돌연변이 DNA의 검출)

  • 조용석;구미자;박귀근;박영서;강종백
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.83-88
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    • 1998
  • To detect the mutation in a given sequence, there are variety of methods developed by use of the gel electrophoresis. One of the methods, TGGE (Temperature Gradient Gel Electrophoresis), is a popular technique because it can detect mutations in DNA fragment with ease and at low cost. This study used 200 bp BamHI-digested DNA fragment containing the human $\varepsilon$-globin promoter which was mutated[$\varepsilon$ F1*(-141), GATA- I*(-163), and GATA-1* & $\varepsilon$F1]. This BamHI-digested DNA fragment was directly used to detect the mutated DNA fragment on 50% denaturant gel with temperature gradient of 45$^{\circ}C$ through $53^{\circ}C$. In agreement with the theoretical result of MELTSCAN program (Brossette and Wallet, 1994) the mobilities of mutated DNA fragments were shown to be nearly distinguished on the temperature gradient gel. In contrast to the above result the heteroduplex analysis under the temperature gradient condition was shown to detect the mutated DNA fragments through the heteroduplex formation between strands of mutated DNA and wild-type DNA.

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Review and application of environmental DNA (eDNA) investigation of terrestrial species in urban ecosystem (도시 내 육상 생물종 모니터링을 위한 환경DNA 리뷰 및 적용)

  • Kim, Whee-Moon;Kim, Seoung-Yeal;Park, Il-Su;Lee, Hyun-Jung;Kim, Kyeong-Tae;Kim, Young;Kim, Hye-Joung;Kwak, Min-Ho;Lim, Tae-Yang;Park, Chan;Song, Won-Kyong
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.23 no.2
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    • pp.69-89
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    • 2020
  • Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.

Use of comet assay as a bioassay in marine organisms exposed to genotoxicants (유전독성물질로 오염된 해양생물의 생물검정법으로서 comet assay 이용)

  • Kim Gi-Beum;An Joon-Gun;Kim Jae-Won
    • Journal of Environmental Science International
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    • v.14 no.11
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    • pp.1071-1079
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    • 2005
  • Using single cell gel electrophoresis, DNA single strand breaks were determined in various marine organisms. DNA damage on fish blood cells was detected to know whether there was a difference between Incheon, Pohang, Masan, and Tongyeong as a control site. Tongyeong showed the lowest DNA damage among the study areas. Mussels, transplanted to Masan Bay for one month, revealed high DNA damage at sites with high economical activity. In two weeks exposure of polychaete to Incheon sediments, higher DNA damage was detected in the sediment adjacent to Incheon harbor than open sea. These results suggested that the marine organism from the polluted area revealed a relatively high DNA damage. In addition, these areas might be contaminated with genotoxic compounds and comet assay was useful as a bioassay to detect DNA damage in marine organisms.