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

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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.

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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Detection of Freshwater Jellyfish (Craspedacusta sowerbii Lankester, 1880) by Biofilm eDNA in Miho River Watershed (미호강 수계 생물막의 환경유전자를 이용한 담수해파리 (Craspedacusta sowerbii Lankester, 1880) 유전자 탐색)

  • Keonhee Kim ;Hyeonjin Cho ;Jeong-Hui Kim;Yun-mo Yang;Hyunji Ju;Hyun-Gi Jeong
    • Korean Journal of Ecology and Environment
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    • v.56 no.3
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    • pp.250-258
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    • 2023
  • Freshwater jellyfish, a type of jellyfish exclusively found in freshwater, has a limited number of species but is found globally. However, their ecology and causes of occurrence are largely unknown. Therefore, understanding the distribution of polyps, which produce the larvae of freshwater jellyfish, can provide important data for comprehending their ecology. This study aims to explore the COI gene of freshwater jellyfish using environmental DNA from the microbial film in the Miho River system. Among the 12 survey points in the Miho River watershed, genetic material of freshwater jellyfish was detected in 8 points, mainly located upstream near reservoirs. These genetic materials were identified as genes of the well-known freshwater jellyfish species, Craspedacusta sowerbii. Notably, the C. sowerbii genes found in the Miho River watershed survey points were closely related to a species previously discovered in Italy. Consequently, utilizing environmental DNA to explore the genetic traces of freshwater jellyfish enables rapid screening of areas with a high likelihood of freshwater jellyfish occurrence. This approach is deemed to provide crucial information for understanding the distribution and ecology of freshwater jellyfish in Korea.

Refinements for the amplification and sequencing of red algal DNA barcode and RedToL phylogenetic markers: a summary of current primers, profiles and strategies

  • Saunders, Gary W.;Moore, Tanya E.
    • ALGAE
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    • v.28 no.1
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    • pp.31-43
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    • 2013
  • This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.

Distribution Status of Invasive Alien Species (Procambarus clarkii (Girard, 1852)) Using Biomonitoring with Environmental DNA in South Korea (생물모니터링 및 환경유전자(eDNA)를 이용한 침입외래종 미국가재(Procambarus clarkii)의 한국분포 현황)

  • Jung, Sang Woo;Lee, Jae-Ha;Kawai, Tadashi;Kim, Phil-Jae;Kim, SuHwan
    • Korean Journal of Environment and Ecology
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    • v.36 no.4
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    • pp.368-380
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    • 2022
  • Biomonitoring of an invasive alien crayfish species, Procambarus clarkii (Girard, 1852), was performed from February to October 2021, along with environmental DNA analysis, at five locations including Wanju-gun, Hampyeong-gun, Naju-si, Gurye-gun, and Cheongju-si. For the investigation, an umbrella-shaped trap for adults and a hand net for young crayfish were used, and 8 to 10L of freshwater was collected for eDNA analysis. The current status and past distributional records of crayfish in Korea were analyzed along with benthic macroinvertebrates at each survey site. As a result of the investigation, a total of 122 individuals were identified, and Hampyeong-gun recorded the largest number of populations with 59 individuals (48.36%) and the highest environmental DNA (eDNA). The frequency of appearance of P. clarkii was highest in May. The ratio of females to males was 21:5, and the body size was 72.2±21.1mm for female, 80.5±15.6mm for male, and 25.3±9.8mm for young crayfish. P. clarkii introduced into Korea is mainly spreading in the southwest region and it has not been observed in Seoul where there had been a record of appearance in the past. No external symbiosis (Branchiobdellida) of P. clarkii has been identified, and P. clarkii that has appeared in Korea was presumed to be imported from Japan. There are more than eight kinds of exotic crayfish distributed in Korea, and among them, the marbled crayfish (P. virginalis) was identified as a harmful species to the ecosystem of Korea by the Ministry of Environment in 2021. The identified species of benthic macroinvertebrates inhabiting the survey area where P. clarkii has appeared were 69 belonging to 39 families, 15 orders, five classes, and three phyla. Among them, Odonata were the most abundant (16 spp.; 24.62%), followed by Coleoptera (11 spp.; 16.92%) and Hemiptera (11 spp.; 16.92%). In the survey area, one Korean endemic species (Rhoenanthus coreanus), one species (Helophorus auriculatus) classified as Near Threatened (NT) on the Korean Red List, and six species of Crustaceans appeared. In the functional feeding group, the predators appeared predominantly while in habitat oriented group, the climbers appeared to be abundant. It was confirmed that P. clarkii in the survey area prefers an area rich in aquatic vegetation with waterside vegetation, and has high resistance to turbid water quality. The omnivore invader P. clarkii is expected to maintain a competitive relationship with carnivorous benthic macroinvertebrates that are predominant in the same species, and is expected to continue to generate ecosystem disturbance along the food chains.

Evaluation of DNA Microarray Approach for Identifying Strain-Specific Genes

  • Hwang, Keum-Ok;Cho, Jae-Chang
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1773-1777
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    • 2006
  • We evaluated the usefulness of DNA microarray as a comparative genomics tool, and tested the validity of the cutoff values for defining absent genes in test genomes. Three genome-sequenced E. coli strains (K-12, EDL933, and CFT073) were subjected to comparative genomic hybridization with DNA microarrays covering almost all ORFs of the reference strain K-12, and the microarray results were compared with the results obtained from in silico analyses of genome sequences. For defining the K-12 ORFs absent in test genomes (reference strain-specific ORFs), we applied and evaluated the cutoff level of -1. The average sequence similarity between ORFs, to which corresponding spots showed a log-ratio of>-1, was $96.9{\pm}4.8$. The numbers of spots showing a log-ratio of <-1 (P<0.05, t-test) were 90 (2.5%) and 417 (10.6%) for the EDL933 genome and the CFT073 genome, respectively. Frequency of false negatives (FN) was ca. 0.2, and the cutoff level of -1.3 was required to achieve the FN of 0.1. The average sequence similarity of the false negative ORFs was $77.8{\pm}14.8$, indicating that the majority of the false negatives were caused by highly divergent genes. We concluded that the microarray is useful for identifying missing or divergent ORFs in closely related prokaryotic genomes.

Effects of Polycyclic Aromatic Hydrocarbons on DNA Damage and Plasma Protein Expression in Mouse

  • Oh, Sang-Nam;Oh, Eun-Ha;Im, Ho-Sub;Jo, Gyu-Chan;Sul, Dong-Geun;Kim, Young-Whan;Lee, Eun-Il
    • Molecular & Cellular Toxicology
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    • v.1 no.1
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    • pp.32-39
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    • 2005
  • Polycyclic aromatic hydrocarbons (PAHs) are an important class of environmentally prevalent xenobiotics that exert complex effects on the biological system and characterized as probably carcinogenic materials. Single cell gel electrophoresis assays were performed in order to evaluate DNA damage occurring in the T-and B lymphocytes, spleens (T/B-cell), bone marrow, and livers of mouse exposed to mixture of PAHs (Benzo(a)pyrene, Benzo(e)pyrene, Fluoranthene, Pyrene) at dose of 400, 800, or 1600 mg/kg body weight for 2 days. DNA damage of the cells purified from mice was increased in dose dependent manner. In the blood cells and organs, DNA damage was also discovered to vary directly with PAHs. Especially T-cells had been damaged more than B-cell. Plasma proteomes were separated by 2-dimensional electrophoresis with pH 4-7 ranges of IPG Dry strips and many proteins showed significant up-and -down expressions with the dose dependent manner. Of these, significant 4 spots were identified using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. Identified proteins were related to energy metabolism and signal transduction.

Identification of Bacteria from Periapical Abscess Using 16S rDNA Clone Libraries. (16S rDNA 클론 Libraries를 이용한 치근단 농양 병소의 세균 동정)

  • 유소영;김미광;김화숙;황호길;김평식;임성훈;오상호;민정범;국중기
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.195-198
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    • 2004
  • Molec-ular analysis was performed on the microflora found In the necrotic pulpal tissue collected from 5 infected root canals that were diagnosed as a periapical abscess. 16S rRNA coding gene (rDNA) library construction and sequencing were performed in order to identify the microflora, The 16S rDNA sequences from 278 clones were identified by a comparison with the database sequence in GenBank. Three phylum and 31 species, which were related to the oral microflora, were identified from the 3 samples (No. 87, 105, and 115). Dialister invisus (5.6%), Peptostreptococcus micron (18.3%), and Veillonella sp. (3.3%) were the organism present in all tee samples. Lac-tobacillusfementum (2.8%),Eubacterumsp./E. infirmum (6.7%), Shuttleworthiasatelles (3.9%), Psudorarnihacfer alactoiyticus (13.3%), Bulleidia moorei (2.8%), and Prevotella denticola (1.1%) were found in two samples. Two phylum and low species of environmental microflora were identified from 2 samples (No.95 and 101). The reason for this might be contamination of the samples with dental water. These results showed that molecular analysis could reveal more diverse microflora that are associated with endodontic infections than that revealed by conventional cultural methods. In addition, these results may of for the basic data to epidemiological studies related with endodontic infection.

Molecular Ecological Stabilities of Genetically Modified 4CB-Degrading Bacteria and Their Gene DNAs in Water Environments (유전공학적으로 변형시킨 4CB 분해세균 및 그 유전자 DNA에 대한 수계에서의 분자생태학적 안정성)

  • Park, Sang-Ho;Myong-Ja Kwak;Ji-Young Kim;Chi-Kyung Kim
    • The Korean Journal of Ecology
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    • v.18 no.1
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    • pp.109-120
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    • 1995
  • As the genetically modified microorganisms (GMMs) and their recombinant plasmid DNAs could be released into natural environments, their stabilities and impacts to indigenous microorganisls have become very importhant research subjects concerning with environmental and ecological aspects. In this study, the genetically modified E. coli CU103 and its recombinant pCU103 plasmid DNA, in which pcbCD genes involving in degradation of biphenyl and 4-chlorobiphenyl were cloned, were studied for their survival and stability in several different waters established under laboratory conditions. E. coli CU103 and its host E. coli XL1-Blue survived longer in sterile distilled water (SDW) and filtered autoclaved river water (FAW) than in filtered river water (FW). A lot of extracellular DNAs were released from E. coli CU103 by lytic action of phages in FW and the released DNAs were degraded by DNase dissolved in the water. Such effects of the factors in FW on stability of the recombinant pCU103 plasmid were also observed in the results of gel electrophoresis, quantitative analysis with bisbenzimide, and transformation assay. Therefore, the recombinant plasmids of pCU103 were found to be readily liberated from the genetically modified E. coli CU103 into waters by normal metabolic processes and lysis of cells. And the plasmid DNAs were quite stable in waters, but their stabilities could be affected by physicoKDICical and biological factors in non-sterile natural waters.

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