• Title/Summary/Keyword: EPSPS

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Production of GMO markers by genetic recombination and their characterization toward immuno-analytical reagents

  • Hwang, Ok-Hwa;Paek, Se-Hwan;Park, Won-Mok
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.220-222
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    • 2003
  • Legislation enacted worldwide to regulate the content of genetically modified organisms (GMOs) in crops, foods, and ingredients, reliable and sensitive methods for GMO detection have been developed. Proteins produced in GMO plants can be determined by qualitative and quantitative analyses and thus GMO designation has performed exactly. Target proteins selected in this study were neomycin phosphotransferase II (NPTII), 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS), cucumber mosaic virus(CMV), and phosphinothricin acetyltransferase (PAT). Analytical method employing western blotting was used for final characterization.

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Effect of an Ethanol Extract of Cassia obtusifolia Seeds on Alcohol-induced Memory Impairment (결명자 에탄올 추출물이 알코올로 유도로 유도한 기억 장애에 미치는 영향)

  • Kwon, Huiyoung;Cho, Eunbi;Jeon, Jieun;Lee, Young Choon;Kim, Dong Hyun
    • Journal of Life Science
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    • v.29 no.5
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    • pp.564-569
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    • 2019
  • Heavy drinking disrupts the nervous system by activation of GABA receptors and inhibition of glutamate receptors, thereby preventing short-term memory formation. Degradation of cognition by alcohol induces blackouts, and it can lead to alcoholic dementia if repeated. Therefore, drugs need to be developed to prevent alcohol-induced blackout. In this study, we confirmed the effect of an ethanol extract of Cassia obtusifolia seeds (COE) on alcohol-induced memory impairment. The effects of COE and ethanol on cognitive functions mice were examined using the passive avoidance and Y-maze tests. The manner in which alcohol affects long-term potentiation (LTP) in relation to the learning and memory was confirmed by electrophysiology performed on mouse hippocampal slices. We also measured N-methyl-D-aspartate (NMDA) receptor-mediated field excitatory synapses (fEPSPs), which have a known association with cognitive impairment caused by ethanol. Ethanol caused memory impairments in passive avoidance and Y-maze tests. COE prevented these ethanol-induced memory impairments in these tests. Ethanol also blocked LTP induction in the mouse hippocampus, and COE prevented this ethanol-induced LTP deficit. Ethanol decreased NMDA receptor-mediated fEPSPs in the mouse hippocampus, and this decrease was prevented by COE. These results suggest that COE might be useful in preventing alcohol-induced neurological dysfunctions, including blackouts.

Reduction of long-term potentiation at Schaffer collateral-CA1 synapses in the rat hippocampus at the acute stage of vestibular compensation

  • Lee, Gyoung Wan;Kim, Jae Hyo;Kim, Min Sun
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.4
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    • pp.423-428
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    • 2017
  • Vestibular compensation is a recovery process from vestibular symptoms over time after unilateral loss of peripheral vestibular end organs. The aim of the present study was to observe time-dependent changes in long-term potentiation (LTP) at Schaffer collateral-CA1 synapses in the CA1 area of the hippocampus during vestibular compensation. The input-output (I/O) relationships of fEPSP amplitudes and LTP induced by theta burst stimulation to Schaffer's collateral commissural fibers were evaluated from the CA1 area of hippocampal slices at 1 day, 1 week, and 1 month after unilateral labyrinthectomy (UL). The I/O relationships of fEPSPs in the CA1 area was significantly reduced within 1 week post-op and then showed a non-significant reduction at 1 month after UL. Compared with sham-operated animals, there was a significant reduction of LTP induction in the hippocampus at 1 day and 1 week after UL. However, LTP induction levels in the CA1 area of the hippocampus also returned to those of sham-operated animals 1 month following UL. These data suggest that unilateral injury of the peripheral vestibular end organs results in a transient deficit in synaptic plasticity in the CA1 hippocampal area at acute stages of vestibular compensation.

Quantification of Genetically Modified Canola GT73 Using TaqMan Real-Time PCR

  • Kim, Jae-Hwan;Song, Hee-Sung;Kim, Dong-Hern;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1778-1783
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    • 2006
  • Event-specific PCR detection methods are the primary trend in genetically modified (GM) plant detection owing to their high specificity based on the flanking sequence of the exogenous integrant. Therefore, this study describes a real-time PCR system for event-specific GM canola GT73, consisting of a set of primers, TaqMan probe, and single target standard plasmid. For the specific detection of GT73 canola, the 3'-integration junction sequence between the host plant DNA and the integrated specific border was targeted. To validate the proposed method, test samples of 0, 1, 3, 5, and 10% GT73 canola were quantified. The method was also assayed with 15 different plants, and no amplification signal was observed in a real-time PCR assay with any of the species tested, other than GT73 canola.

CORRELATIONS BETWEEN HIPPOCAMPAL THETA RHYTHM AND INTRACELLULAR CHARACTERISTICS OF PYRAMIDAL NEURONS (해마 theta 리듬과 pyramidal neuron의 세포내 특성과의 상관관계)

  • Kwon, Oh-Heung;Kim, Young-Jin;Nam, Soon-Hyeun;Kim, Hyeun-Jung;Lee, Man-Gee;Cho, Jin-Hwa;Choi, Byung-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.25 no.4
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    • pp.671-682
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    • 1998
  • Electrophysiological phenomena of pyramidal cells in the CA1 area of the dorsal hippocampus were recorded from and filled with neurobiotin in anesthetized rats. The electropharmacological properties of membrane as well as the cellular-synaptic generation of rhythmic slow activity (theta) were examined. The intracellular response characteristics of these pyramidal cells were distinctly different from responses of interneurons. Pyramidal cells had a high resting membrane potential, a low input resistance, and a large amplitude action potential. A afterhyperpolarization was followed a single action potential. Most of pyramidal cells did not display a spontaneous firing. Pyramidal cells displayed weak inward rectification and anodal break excitation. The slope of the frequency-current relation was 53.4 Hz/nA for the first interspike interval and 15.9 Hz/nA for the last intervals, suggesting the presence of spike frequency adaptation. Neurobiotin-filled neurons showed pyramidal morphology. Cells were generally bipolar dendritc processes ramifying in stratum lacunosum-moleculare, radiatum, and oriens. Commissural stimulation discharged pyramidal cells, followed by excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs). The frequency of theta-related membrane potential oscillation was voltage-independent in pyramidal neurons. At strong depolarization levels (less than 30 mV) pyramidal cells emitted sodium spike oscillation, phase-locked to theta. The observations provide direct evidence that theta-related rhythmic hyperpolarization of principal cells is brought by the rhythmically discharging interneurons. Furthermore, the findings in which interneurons were also paced by rhythmic inhibitory postsynaptic potentials during theta suggest that they were periodically hyperpolarized by their GABAergic septal afferents.

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Effects of Fermented Scutellaria Baicalensis Extract on H2O2 - Induced Impairment of Long-term Potentiation in Hippocampal CA1 Area of Rats (흰쥐 해마 CA1 영역에서 H2O2에 의한 장기강화 억제에 대한 발효황금 추출물의 효과)

  • Heo, Jun Ho;Rong, Zhang Xiao;Kim, Min Sun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.6
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    • pp.356-362
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    • 2019
  • Scutellaria baicalensis (SB) has widely used in the treatment for various brain diseases in the field of Oriental medicine. Biofermantation of SB can make major chemical constituents of SB to pass blood-brain barrier easily and to have more potent anti-oxidant ability. There is a little information about the contribution of fermented SB (FSB) to the formation or maintenance of the neural plasticity in the hippocampus. The purpose of this study was to evaluate effects of FSB extract on hydrogen peroxide (H2O2) - induced impairments of the induction and maintenance of long-term potentiation (LTP), an electrophysiological marker for the neural plasticity in the hippocampus. From hippocampal slices of rats, the field excitatory postsynaptic potentials (fEPSPs) were evoked by the electrical stimulation to the Schaffer collaterals - commissural fibers in the CA1 areas and LTP by theta-burst stimulation by using 64 - channels in vitro multi-extracellular recording system. In order to induce oxidative stress to hippocampal slices two different concentrations (200, 400 μM) of H2O2 were given to the perfused aCSF before and after the LTP induction, respectively. The ethanol extract of FBS with concentration of 25 ㎍/ml, 50 ㎍/ml was diluted in perfused aCSF that had 200 μM H2O2, respectively. Oxidative stress by the treatment of H2O2 resulted in decrease of the induction rate of LTP in the CA1 area with a dose - dependent manner. However, the ethanol extract of FSB prevented the reduction of the induction rate of LTP caused by H2O2 - induced oxidative stress with a dose - dependent manner. These results may support a potential application of FSB to ameliorate impairments of hippocampal dependent neural plasticity or memory caused by oxidative stress.

Impact of a Glyphosate-Tolerant Soybean Line on the Rhizobacteria, Revealed by Illumina MiSeq

  • Lu, Gui-Hua;Zhu, Yin-Ling;Kong, Ling-Ru;Cheng, Jing;Tang, Cheng-Yi;Hua, Xiao-Mei;Meng, Fan-Fan;Pang, Yan-Jun;Yang, Rong-Wu;Qi, Jin-Liang;Yang, Yong-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.561-572
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    • 2017
  • The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.

An Improved Method to Determine Corn (Zea mays L.) Plant Response to Glyphosate (Glyphosate에 대한 옥수수 반응의 개선된 검정방법)

  • Kim, Jin-Seog;Lee, Byung-Hoi;Kim, So-Hee;Min, Suk-Ki;Choi, Jung-Sup
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.57-62
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    • 2006
  • Several methods for determining the response of corn to glyphosate were investigated to provide a fast and reliable method for identifying glyphosate-resistant corn in vivo. Two bioassays were developed. One assay is named 'whole plant / leaf growth assay', in which the herbicide glyphosate is applied on the upper part of 3rd leaf and the growth of herbicide-untreated 4th leaf is measured at 3 day after treatment. in this assay, the leaf growth of conventional corn was inhibited in a dose dependent from 50 to $1600{\mu}g/mL$ of glyphosate and growth inhibition at $1600{\mu}g/mL$ was 55% of untreated control. The assay has the potential to be used especially in the case that the primary cause of glyphosate resistance is related with a reduction of the herbicide translocation. Another assay is named 'leaf segment / shikimate accumulation assay', in which the four excised leaf segments ($4{\times}4mm$) are placed in each well of a 48-well microtiter plate containing $200{\mu}L$ test solution and the amount of shikimate is determined after incubation for 24 h in continuous light at $25^{\circ}C$. In this assay, 0.33% sucrose added to basic test solution enhanced a shikimate accumulation by 3 to 4 times and the shikimate accumulation was linearly occurred from 2 to $8{\mu}g/mL$ of glyphosate, showing an improved response to the method described by Shaner et al. (2005). The leaf segment / shikimate accumulation assay is simple and robust and has the potential to be used as a high throughput assay in the case that the primary cause of glyphosate resistance is related with EPSPS, target site of the herbicide. Taken together, these two assays would be highly useful to initially select the lines obtained after transformation, to investigate the migration of glyphosate-resistant gene into other weeds and to detect a weedy glyphosate-resistant corn in field.