• Title/Summary/Keyword: Environmental DNA

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Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

Comparative Study of DNA Extraction Method in Meiofauna (중형저서동물에서 효율적인 DNA 추출 방법 비교 연구)

  • Lee, Seung-Han;Back, Jin-Wook;Lee, Won-Choel
    • Korean Journal of Environmental Biology
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    • v.29 no.3
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    • pp.138-143
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    • 2011
  • The efficiency of mtCOI amplication after DNA extraction of benthic harpacticoid Tigriopus japonicus s.l. was tested under different conditions depending on fixative (99% Ethanol, or 4% Formalin) and additional chemicals (Ludox or Rose Bengal). Each experimental group by the fixative was subdivided into four groups, respectively: 1) Control (fixative only), 2) processed with Ludox HS40, 3) processed with Rose Bengal, and 4) processed with both Ludox HS40 and Rose Bengal. For the 99% ethanol-fixed sample, overall success rate of amplification by PCR was 96% or above, while for the 4% formalin-fixed one, success rate was much lower than those of ethanol-fixed: 1) Control: 27%, 2) Ludox HS40: 3%, 3) Rose Bengal: 7%, and 4) Ludox HS40 and Rose Bengal: 3%. As a result present study verify that 99% ethanol is a proper fixative for DNA extraction in meiofauna organisms.

Electrochemical Detection of $17{\beta}-estradiol$ by using DNA Aptamer Immobilized Nanowell Gold Electrodes

  • Kim, Yeon-Seok;Jung, Ho-Sup;Lee, Hea-Yeon;Kawai, Tomoji;Gu, Man-Bock
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.88-92
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    • 2005
  • Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

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Biological Characteristics of Anodic Electrolyzed Water (산성전리수의 생물학적 특성)

  • 김윤경;민병술;민중기;이종권;이윤배;류근걸;이미영
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.265-272
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    • 2004
  • Biological characteristics of anodic electrolyzed water were investigated in this study. Linear DNAs which were incubated at $4^\circ{C}$ and $25^\circ{C}$ for 10 mins in the anodic electrolyzed water were degraded about 40% and 50%, respectively. But the DNA was amplified pretty well without any degradation through polymerase chain reaction in the presence of anodic electrolyzed water. Protein degradation hardly occurred in the distilled water during entire incubation time of 7 days, while protein began to be degraded from 4 days in the anodic electrolyzed water. Rice seeds could germinate in the distilled water and anodic electrolyzed water with the same germination ratio, however, the anodic electrolyzed water inhibited the growth of roots and total length of rice seedlings in the soil. Anodic electrolyzed water did not affect the growth curve and cell number of marine alga significantly. The anodic electrolyzed water inhibited the browning of potato by inactivating 50% of polyphenol oxidase activity.

Genetic Diversity of Rana amurensis (Amphibia: Ranidae), Based on Mitochondrial 165 rDNA Gene Sequences (미토콘드리아 16S rDNA를 이용한 아무르산개구리 (양서 강: 개구리 과)의 유전적 다양성)

  • 송재영;윤병수;오홍식;정규회
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.45-51
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    • 2003
  • Genetic diversity of local populations among geologically isolated groups of Rana amurensis was refined by sequence comparison of the mitochondrial (mt) 165 rDNA genes. Each 401 base pairs of DNA sequences, which was determined from four local populations of Rana amurensis, two local populations of R. nigromacutata, and three species of the genus Rana were used in this analysis. Despite morphological similarity of Rana amurensis, Korean populations were well distinguished from the other groups on the basis of 105 rDNA gene difference. Further analyses for additional local populations belonging to R. amurensis will be necessary to clarify the taxonomic status.

Allomyrina Dichotoma Larva Extracts Protect Streptozotocin-induced Oxidative Cytotoxicity (Allomyrina Dichotoma Larva 추출물이 췌장 ${\beta}$-세포에서 streptozotocin에 의해 유도된 산화적 손상에 대한 보호효과)

  • Kim, Deok-Song;Huh, Jin;You, Guen-Chang;Chae, Soo-Chul;Lee, Oh-Sun;Lee, Hwang-Hee Blaise;Lee, Jong-Bin;Kim, Jong-Sun
    • Environmental Analysis Health and Toxicology
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    • v.22 no.4
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    • pp.349-355
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    • 2007
  • 장수풍뎅이 유충(Allomyrina dichotoma larva, ADL)은 중국의 전통 약재로서, 특히 항산화 효과가 우수하여 항당료 제재로 사용되고 있다. 본 연구에서는 이러한 ADL의 추출물을 이용하여 헴스터 췌장의 ${\beta}$-세포(HIT-T15)에서 Streptozotocin에 의해 유발된 산화적 손상에 대한 보호효과 및 그 작용기전을 조사하였다. ADL추출물은 처리농도 의존적으로 Streptozotocin에 의해 유발된 지질과산화 및 세포 내 자유산 소종의 양을 억제함으로서 ${\beta}$-세포의 산화적 스트레스에 의한 손상을 보호하였다. 또한 DNA laddering 방법을 사용하여 Streptozotocin에 의해 유발된 DNA 손상을 조사한 결과, ADL추출물 처리농도에 비례하여 Streptozotocin에 의해 유발된 DNA 손상이 감소하였다. 이러한 산화적 손상의 억제능 관련 작용 기전을 조사하기 위해 DPPH free radical 소거능을 실시하였다. 그 결과 ADL추출물 자체가 DPPH 자유 레디컬 소거능이 있음을 확인하였으며, 또한 플라스미드를 이용한 Single-strand break 방법을 통한 DNA 손상 보호능을 측정한 결과도 $Fe^{3+}$$H_2O_2$에 의해 유발된 DNA 손상이 ADL추출물 처리농도에 비례하여 감소하였다. 이러한 결과들을 종합하여 볼 때, 장수풍뎅이 유충의 추출물들이 자체의 레디컬 소거능 및 산화적 손상에 의한 DNA손상을 억제함으로써, Streptozotocin에 의해 유발된 산화적 손상을 억제할 수 있을 것이라 사료된다.

Mechanism study on DNA damage and Apoptosis induced by heak shock using Comet Assay

  • Seo, Young-Rok;Han, Sung-Sik;Kim, L. O′Neill;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 1997.12a
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    • pp.101-101
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    • 1997
  • Comet assay, single cell gel electrophoresis has been known as useful, rapid, simple, visual, and sensitive technique for measuring the DNA breakage in mammalian ce1ls. For evaluation of DNA damage using comet assay, early studies reported a change in comet length and intensity with DNA damage using simple visual technique, such as fluorescence microscopy with eyespiece. In recent, some workers are observing and analyzing nucleotide of comets using quantitative fluorescence image analysis system to estimate 'tail moment', which is defined as the product of the tail length and the fraction of total DNA in tail. Our laboratory also adopted the image analysis software for qualification. In addition, many of the practical features of comet assay render it potentially attractive as useful tool for molecular toxicology and carcinogenesis, because the system is already showing considerable promise as rapid predictor in both in vitro and in vivo experimental designs. Recently, the comet assay becomes a attractive technique to study of apoptosis, because apoptotic fragmentation of nuclear DNA into nucleosomal sizes can be evaluated by the comet assay. So, we attempted to apply the comet assay to studying the effect of various stress on the apoptosis-sensitive cell lines. Particularly, focusing on the hyperthermic apoptosis, we could find that heat shock(44˚C for 60 minutes) was sufficient to induced apoptosis in these cell lines. But using the highly sensitive comet assay, we could not detect DNA breaks immediately after heat shock.

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Comet Assay to Detect the DNA Breakages in the Tissue of the Purple Clam ( Saxidomus purpuratus) and the Blood of the Olive Flounder (Paralichthys olivaceus) Exposed to 5 PAHs

  • Lee, Taek-Kyun;Kim, So-Jung;Park, Eun-Seok;Rora Oh;Yun, Hee-Young;Man Chang
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.159-159
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    • 2003
  • Comet assay is a potential monitoring tool because DNA strand breakage may be produced by a wide range of agents. The comet assay, also called the single-cell gell electrophoresis (SCGE) assay, is rapid and sensitive method for the detection of DNA damage in cells. This study was performed for the identification of DNA damage in the cells from flounders and clams exposed to PAHs. As a control experiments, flounder and clam cells were exposed to $H_2O$$_2$. The cells exposed to $H_2O$$_2$ were displayed a typical nuclei movement DNA damage of cells were significantly increased when the isolated cells from the blood of flounders and the tissue of clams were in vitro exposed to the different concentrations (5, 10, 50, 100 ppb) of five kinds of PAHs (benzo[a]pyrene, pyrene, fluoranthene, anthrancene, and phenanthrene). For the in vivo test, flounders and clams were exposed to the different concentrations of BaP for 4 days. The results showed that DNA strand breakage was effected by the concentration of BaP and the duration of exposure. In high concentration of BaP, the mean tail lengths of nuclei was longer than it In low concentration, while the mean size of head DNA decreased. In this research, both in vitro and in vivo genotoxicity of PAHs could be biomonitored by the comet assay. Especially, clams and flounders seem to be useful as materials for monitoring genotoxic damage by comet assay.

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