• Title/Summary/Keyword: living modified organisms

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Intraspecific Androgenesis in Mud Loach (Misgurnus mizolepis): II. Diploid Restoration and Viability Assessment (미꾸라지(Misgurnus mizolepis)의 웅성발생성 처녀생식: II. 웅성발생성 이배체 유도 및 생존능력 평가)

  • Nam, Yoon-Kwon;Bang, In-Chul;Noh, Choong-Hwan;Cho, Young-Sun;Kim, Dong-Soo
    • Journal of Aquaculture
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    • v.19 no.4
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    • pp.281-287
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    • 2006
  • Intraspecific diploid androgenesis was achieved in mud loach (Misgurnus mizolepis) by the inhibition of the first mitotic division using combined thermal treatment. A combined thermal treatment (heat shock at $40.5\;^{\circ}C$ for 120 sec followed by cold treatment at $1\;^{\circ}C$ for 45 min) applied to the 1st metaphase of cell division (28 min post insemination at $25\;^{\circ}C$) successfully recovered viable androgenetic diploidy. Mean hatching success of the androgenetic diploid group was 29.6%, and the average yield out of total eggs taken was about 7% assessed at 1 week of age. However, relatively large variations in the yield of diploid androgenesis were observed among different egg batches used as cytoplasmic donors. Successful diploidization was confirmed by flow cytometric analysis, and parthenogenic reproduction in a paternal exclusive manner was verified with transgene dosage. Significant mortality was found in most androgenetic groups especially from hatch to 1 month of age, although such mortality was stabilized later.

Cytogenetic Analysis of Korean Shinner, Coreoleuciscus splendidus (Cyprinidae) (쉬리, Coreoleuciscus splendidus (Cyprinidae)의 세포유전학적 연구)

  • Kim, Dong-Soo;Song, Ha-Yeun;Bang, In-Chul;Nam, Yoon-Kwon
    • Journal of Aquaculture
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    • v.20 no.2
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    • pp.140-143
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    • 2007
  • Cytogenetic analyses of an endemic species, Coreoleuciscus splendidus (Cyprinidae) was performed including erythrocyte measurement, chromosome count and karyotyping, nucleolar organizing region (NOR) banding and flow cytometric analysis of genome size. C. splendidus had the same modal chromosome number of 2n = 48 between sexes, however, displayed a sex-related dimorphism in their chromosome karyotypes. Males represented a pair of heteromorphic chromosomes which couldn‘t be seen in any female individuals, indicating that the sex determination mechanism of this species should be a typical XX-XY based male heterogamety (female=10M+6SM+8A+XX vs male=10M+6SM+8A+XY). Other cytogenetic features such as Ag-NORs located in a pair of acrocentric chromosomes, estimated nuclear volume ($28{\mu}m^3$) and cellular DNA content (2.4 pg/cell) suggest that genetic recombination might be the main driving force responsible for the evolution of this species rather than the polyploidy-based evolutionary process as in many other Cyprinidae species.

Monoclonal antibody production for CP4 EPSPS detection assays (CP4 EPSPS 검출을 위한 단클론 항체 생산)

  • A-Mi Yoon;Il Ryong Kim;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.445-451
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    • 2021
  • In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

Simulative Study of Effects of LM Microorganism on Environment: Analyses of Metabolomes and Soil Microbial Communities (LM 미생물의 환경영향 모사: 대사체와 토양미생물군 분석)

  • Lee, Ji-Hoon;Ki, Min-Gyu
    • Korean Journal of Environmental Agriculture
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    • v.38 no.3
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    • pp.197-204
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    • 2019
  • BACKGROUND: Living modified microorganisms (LMMs) have been focused in two very different aspects of positive and negative effects on ecology and human health. As a model experiment, wild type and a foreign origin gene-harboring modified E. coli strains were subjected to comparison of their metabolomes and potential effects on soil microbiota in the laboratory sets. This study assumes the unintentional release of LMMs and tries to suggest potential effects on the soil microbiota even at minimal settings. METHODS AND RESULTS: Metabolomes from the wild type and LM E. coli were analyzed by NMR and the profiles were compared. In the laboratory soil experiments, the two types of E. coli were added to the soils and monitored for the bacterial community compositions. Those metabolomic profiles did not show significant differences. The microbial community structures from the time series soil DNAs for both the sets using wild type and LMO also did not indicate significant changes, but minor by the addition of foreign organisms regardless of wild or LMO. CONCLUSION: Even if the recombinant microorganism (LMO) is released into the soil environment, the survival of microorganisms in the environment would be one of the major factors for the transfers of foreign genes to other organisms and diffusion into the soil environment.

Early Gonadogenesis and Sex Differentiation of a Marine Medaka, Oryzias dancena (Beloniformes; Teleostei) (해산 송사리 Oryzias dancena의 초기 생식소 형성 및 성분화)

  • Song, Ha Yeun;Nam, Yoon Kwon;Bang, In-Chul;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.21 no.3
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    • pp.141-148
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    • 2009
  • Sex differentiation and gonad development were investigated in a marine medaka species, Oryzias dancena (Beloniformes; Teleostei). The average time to hatch was 11 days post-fertilization (dpf) at $25^{\circ}C$. Primordial germ cell (PGC) was first observed at 5 dpf and migrated to presumptive gonadal area between the gut and pronephric duct at 9 dpf. Male and female gonads were morphologically differentiated at 12 days post-hatching (dph). Early oocytes at perinucleolus stage as well as the formation of spermatid and efferent duct were observed at 28 dph. At 6 weeks of age, the ovary exhibited yolk granulation in many oocytes, while testis possessed a considerable number of spermatogonia and spermatids. The first ovulation was observed in 9-week-old females, and at the same age, males contained fully-matured spermatozoa. Data obtained in this study indicate that the gonad differentiation of O. dancena is the typical type of differentiated gonochorism.

Embryogenesis and Early Ontogenesis of a Marine Medaka, Oryzias dancena (바다 송사리 Oryzias dancena의 난발생 및 자치어의 형태 발달)

  • Song, Ha Yeun;Nam, Yoon Kwon;Bang, In-Chul;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.21 no.4
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    • pp.227-238
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    • 2009
  • The egg development and morphological changes of larvae, juveniles and adults of Oryzias dancena were observed. Fertilized eggs were incubated at $25{\pm}1^{\circ}C$; the process of embryonic development was observed by light microscopy and based on diagnostic features of the developing embryos. The average time to hatch was 11 days after fertilization. The hatched larvae averaged $4.40{\pm}0.24mm$ in total length (TL). The yolk sacs of the larvae were almost absorbed at 3 days after hatching and $4.55{\pm}0.23mm$ TL. At 21 days after hatching, the larvae were $7.23{\pm}0.73mm$ TL and had reached the juvenile stage. First ovulation was about 9 weeks after hatching and at $22.58{\pm}2.73mm$ TL.

Hepatic Expression of Cu/Zn-Superoxide Dismutase Transcripts in Response to Acute Metal Exposure and Heat Stress in Hemibarbus mylodon (Teleostei: Cypriniformes)

  • Cho, Young-Sun;Bang, In-Chul;Lee, Il-Ro;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.179-184
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    • 2009
  • Hemibarbus mylodon (Cypriniformes) is an endemic freshwater fish species in the Korean peninsula, for which urgent conservation efforts are needed. To understand their stress responses in relation to metal toxicity and thermal elevation, we performed a real-time RT-PCR-based expression assay of hepatic copper/zinc-superoxide dismutase (Cu/Zn-SOD), a key antioxidant enzyme, in response to experimental heavy metal exposure or heat treatment. The transcription of hepatic Cu/Zn-SOD was differentially modulated by acute exposure to Cu, cadmium (Cd), or Zn. Exposure to each metal at $5{\mu}M$ for 24 h revealed that Cu stimulated the mRNA expression of Cu/Zn-SOD to a greater extent than the other two heavy metals. The elevation in Cu/Zn-SOD transcripts in response to Cu exposure was dose-dependent (0.5 to $5{\mu}M$). Time course analysis of Cu/Zn-SOD expression in response to Cd exposure ($5{\mu}M$) revealed a transient pattern up to day 7. Exposure to thermal stress (an increase from 22 to $30^{\circ}C$ at a rate of $1^{\circ}C/h$ followed by $30^{\circ}C$ for 18 h) did not significantly alter SOD transcription, although heat shock protein 90 kDa (HSP90) transcription was positively correlated with an increase in temperature.