• Title/Summary/Keyword: Environmental DNA

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T-RFLP Analysis of Microbial Community Structure in Leachate from Landfill Sites (폐기물매립장 침출수내 미생물군집 구조 해석을 위한 T-RFLP의 활용)

  • Yu, Jae-Cheul;Ishigaki, Tomonori;Kamagata, Yoichi;Lee, Tae-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.4
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    • pp.369-378
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    • 2010
  • Microorganisms are key-role player for stabilization of landfill sites. In order to evaluate the availability of T-RFLP(Terminal Restriction Fragment Length Polymorphism) for monitoring microbial community variations during stabilization of landfill sites, the phylogenic diversity of microbial community in the leachate from 4 different full-scale landfills was characterized by T-RFLP based on bacterial 16S rDNA. Main population of microbial community analyzed by T-RFLP was significantly similar with that of microbial community analyzed by clone library analysis. The results of T-RFLP analysis for main population of microbial community in the leachate from landfills with different landfill structures, waste types and landfill ages showed apparently different microbial diversity and structures. Therefore, long-term monitoring of microbial community in leachate from landfill sites by using T-RFLP is expected to be available for evaluation of landfill stability.

Molecular Characterization and Infectious cDNA Clone of a Korean Isolate of Pepper mild mottle virus from Pepper

  • Yoon, Ju-Yeon;Hong, Jin-Sung;Kim, Min-Jea;Ha, Ju-Hee;Choi, Gug-Seon;Choi, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.361-368
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    • 2005
  • A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased hot pepper plant and its biological and molecular properties were compared to that of PMMoV-J and PMMo V -So The genomic RNA of PMMoV-Kr consists of 6,356 nucleotides. The nucleotide and amino acid sequences identities of four viral proteins and two noncoding regions among PMMoV-Kr, PMMoV-S and PMMoV-J were $96.9\%\;to\;100.0\%\;and\;97.5\%\;to\;98.6\%$, respectively. Full-length cDNA amplicon of PMMoV-Kr was directly amplified by RT-PCR with a set of 5'-end primer anchoring T7 RNA promoter sequence and 3'-end virus-specific primer. Capped transcript RNAs from the full-length cDNA clone were highly infectious and caused characteristic symptoms of wild type PMMoV when mechanically inoculated to systemic host plants such as Nicotiana benthamiana and pepper plants.

Genome-Based Reclassification of Strain KIST612, Previously Classified as Eubacterium limosum, into a New Strain of Eubacterium callanderi

  • Ji-Yeon Kim;Byeongchan Kang;Soyoung Oh;Yeji Gil;In-Geol Choi;In Seop Chang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.8
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    • pp.1084-1090
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    • 2023
  • The strain KIST612, initially identified as E. limosum, was a suspected member of E. callanderi due to differences in phenotype, genotype, and average nucleotide identity (ANI). Here, we found that E. limosum ATCC 8486T and KIST612 are genetically different in their central metabolic pathways, such as that of carbon metabolism. Although 16S rDNA sequencing of KIST612 revealed high identity with E. limosum ATCC 8486T (99.2%) and E. callanderi DSM 3662T (99.8%), phylogenetic analysis of housekeeping genes and genome metrics clearly indicated that KIST612 belongs to E. callanderi. The phylogenies showed that KIST612 is closer to E. callanderi DSM 3662T than to E. limosum ATCC 8486T. The ANI between KIST612 and E. callanderi DSM 3662T was 99.8%, which was above the species cut-off of 96%, Meanwhile, the ANI value with E. limosum ATCC 8486T was not significant, showing only 94.6%. The digital DNA-DNA hybridization (dDDH) results also supported the ANI values. The dDDH between KIST612 and E. callanderi DSM 3662T was 98.4%, whereas between KIST612 and E. limosum ATCC 8486T , it was 57.8%, which is lower than the species cut-off of 70%. Based on these findings, we propose the reclassification of E. limosum KIST612 as E. callanderi KIST612.

Evaluation of Nonanchored Inter Simple Sequence Repeat (ISSR) Marker to Detect DNA Damage in Common Bean (Phaseolus vulgaris L.) Exposed to Acrylamide

  • Enan, Mohamed R.
    • Journal of Forest and Environmental Science
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    • v.24 no.2
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    • pp.61-68
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    • 2008
  • Acrylamide is present as a contaminant in heated food products, predominantly from the precursor asparagine. Nonanchored inter simple sequence repeats (ISSRs) are arbitrary multiloci markers produced by PCR amplification with a microsatellite primer. In order to assess the feasibility of microsatellite primers as markers for DNA damage, the study was conducted on common bean (Phaseolus vulgaris L.) exposed to different concentrations of acrylamide. Polymorphisms were abundant among plant samples treated with acrylamide in comparison to control (untreated one) tested with 4- tri-nucleotide, 2 tetra-nucleotide, and 3- dinucelotide primers. The primer (CCG)4 was the best tested primer to generate polymorphism between the DNA of plants treated or not by acrylamide. Polymorphisms became evident as the presence and absence of DNA fragments in treated samples compared with the untreated one. The highest number of DNA variation on ISSR patterns was observed at the micromollar concentrations of acrylamide. Acrylamide was able to induce DNA damage in non concentration-dependent manner with effectiveness at micromollar concentrations. This study demonstrated that ISSR markers can be highly reliable for identification of DNA damage induced by acrylamide.

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Chronological Switch from Translesion Synthesis to Homology-Dependent Gap Repair In Vivo

  • Fujii, Shingo;Isogawa, Asako;Fuchs, Robert P.
    • Toxicological Research
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    • v.34 no.4
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    • pp.297-302
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    • 2018
  • Cells are constantly exposed to endogenous and exogenous chemical and physical agents that damage their genome by forming DNA lesions. These lesions interfere with the normal functions of DNA such as transcription and replication, and need to be either repaired or tolerated. DNA lesions are accurately removed via various repair pathways. In contrast, tolerance mechanisms do not remove lesions but only allow replication to proceed despite the presence of unrepaired lesions. Cells possess two major tolerance strategies, namely translesion synthesis (TLS), which is an error-prone strategy and an accurate strategy based on homologous recombination (homology-dependent gap repair [HDGR]). Thus, the mutation frequency reflects the relative extent to which the two tolerance pathways operate in vivo. In the present paper, we review the present understanding of the mechanisms of TLS and HDGR and propose a novel and comprehensive view of the way both strategies interact and are regulated in vivo.

GENE-SPECIFIC OXIDATIVE DNA DAMAGE IN HELICOBACTER PYLORI INFECTED HUMAN GASTRIC MUCOSA

  • Jinhee Chol;Yoon, Sun-Hee;Kim, Ja-Eun;Rhee, Kwang-Ho;Youn, Hee-Sang;Chung, Myung-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05a
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    • pp.84-84
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    • 2002
  • Abstract To study the status of oxidative DNA damage in Helicobacter pylori infection in more details, gene-specific oxidative DNA damage was investigated by examining oxidative DNA damage to individual genes. This was done by determining the loss of PCR product of a targeted gene before and after gastric mucosal DNA was treated with 8-hydroxyguanine glycosylase, which cleaves DNA at the 8-hydroxyguanine residues.(omitted)

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Characterization of Excision Repair Genes Related to Damaged DNA Repair from Eukaryotic Cells

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.17 no.1
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    • pp.1-6
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    • 1997
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

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Antigenotoxicity and Action Mechanism of Quercetin and its Glycosides against Oxidative DNA Damage (Oxidative DNA 손상에 대한 Quercetin 및 그 배당체들의 유전독성억제효과와 작용기전)

  • 김수희;허문영
    • Environmental Mutagens and Carcinogens
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    • v.19 no.2
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    • pp.116-121
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    • 1999
  • Quercetin and its glycosides showed a strong free radical scavenging effect to DPPH radical generation. However, there were not big differences between quercetin aglycone and glycosides under experimental condition of this study. On the other hand, quercetin had pro-oxidant effect in bleomycin-dependent DNA assay. Quercetin aglycone and its glycosides, quercitrin inhibited $H_2$$O_2$- induced DNA damage in CHL cells. They also have an anticlastogenicity toward DNA breakage agent by radical generation like bleomycin. These results indicate that quercetin aglycone and its glycosides are capable of protecting the free radical generation induced by reactive oxygen species like $H_2$$O_2$. The mechanism of inhibition in hydrogen peroxide-induced genotoxicity may be due to their free radical scavenging properties. Therefore, quercetin aglycone and its glycosides may be useful chemopreventive agents by protecting of free radical generation which are involved in carcinogenesis and aging. However, quercetin and its glycosides must also carefully examined for pro-oxidant properties before being proposed for use in vivo.

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Diversity and its Characteristics of Life Phenomenon (생명현상의 다양성과 특징에 관한 고찰 - 보건의료에 미치는 영향을 중심으로 -)

  • Lee, Sun-Dong
    • Journal of Society of Preventive Korean Medicine
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    • v.14 no.1
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    • pp.1-12
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    • 2010
  • This research was intended to delve into the diversity of life phenomenon and its characteristics. First of all, this research gave real examples to compare the differences in men's health, disease, and longevity in order to confirm the existence of diversity of life phenomenon. In addition, it also studied the process and mechanism of manifestation of life phenomenon, as well as the influence and problems of existing studies' results and implications. The results are as follow. 1. Differences in health, diseases, and longevity were very big and diverse in researches on different races, nations, ages, socioeconomic status, positions, and even (monozygotic) twins. 2. The basic foundation of all organisms is DNA, and environmental factors change DNA methylation and the structure of chromatin by constantly influencing DNA. Due to this, the manifestation, control, and phenotype of DNA change, resulting in diversified life phenomenon. Therefore, it is the environmental factors, not DNA, that has more influence on the diversity of life. 3. Looking at available studies, the most reasonable perspective on human requires focusing on the diversity of life phenomenon, holistic thinking, and reversible change instead of irreversible determinism. Considerable differences in life phenomenon between entities require a change in malformed perspective on life. Public health and medicine deals with live human beings, a more precise and accurate perspective on life is very important. Because management methods of health and disease, such as structure and approaches of medical research, prevention and cure, must be different by life perspectives.

A New Record of Euonymus hamiltonianus Wall. in Dokdo: Using Molecular Markers of Nuclear and Plastid DNA (독도의 미기록 식물 참빗살나무: 핵과 엽록체 DNA의 분자마커 이용)

  • Song, Im-Geun;Park, Seon-Joo
    • Korean Journal of Environmental Biology
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    • v.32 no.1
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    • pp.88-94
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    • 2014
  • The present scientific scenario shows that, the long-distance dispersal of any species plays a major role in population as well as the flow of individual between populations. However, newly recorded plant at Dokdo was confirmed by using morphological and molecular markers of nrDNA and cpDNA. This scientific study has identified the plant species was as a Euonymus hamiltonianus Wall. (Celastraceae). And after careful studies, the laboratory results shows that the distribution of tree species at Dokdo as an Oceanic Island is a meaningful thing because of the inflow of plant in adverse condition of environment such as a shallow soil layers and steep terrain. So it is necessary to continuous monitor for the inflow of non-indigenous plant into Dokdo.