• Proceedings of the PSK Conference
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• 2002.04a
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• pp.141.2-142
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• 2002

• KSBB Journal
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• v.32 no.2
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• pp.90-95
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• 2017

Mammalian cell cultures have been used extensively to produce proteins for therapeutic agent because of their ability to perform post-translational modification including glycosylation. To produce recombinant protein, many factors and parameter are considered such as media composition, host cell type, and culture process. In this study, recombinant human erythropoietin (rhEPO) producing cell line was established by using glutamine synthetase system. To reduce serum concentration in media, we compared direct adaptation with step adaptation. Cell growth was faster in step adaptation. In low-level serum media, there were insufficient glucose for cell growth. Thus, we added glucose in low-level serum media from 2 g/L to 4.5 g/L. Titer of rhEPO was higher than other conditions at 4.5 g/L of glucose. Additionally, N-methyl-D-aspartate (NMDA), 13-cis-retinal, and pluronic F-68 (PF-68) were added to enhance productivity in CHO cell cultures. In conclusion, we applied CHO cell producing rhEPO to low-level of serum in media using step-adaptation. Also, we confirmed positive effect of NMDA, 13-cis-retinal, and PF-68.

• The Plant Pathology Journal
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• v.31 no.2
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• pp.115-122
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• 2015

Anthracnose, caused by Colletotrichum gloeosporioides (C. gloeosporioides; Teleomorph: Glomerella cingulata), is the most destructive disease that affects sweet persimmon production worldwide. However, the biology, ecology, and genetic variations of C. gloeosporioides remain largely unknown. Therefore, in this study, the development of fungicide resistance and genetic diversity among an anthracnose pathogen population with different geographical origins and the exposure of this population to different cultivation strategies were investigated. A total of 150 pathogen isolates were tested in fungicide sensitivity assays. Five of the tested fungicides suppressed mycelial pathogen growth effectively. However, there were significant differences in the sensitivities exhibited by the pathogen isolates examined. Interestingly, the isolates obtained from practical management orchards versus organic cultivation orchards showed no differences in sensitivity to the same fungicide. PCR-restriction fragment length polymorphism (RFLP) analyses were performed to detect internal transcribed spacer regions and the ${\beta}$-tubulin and glutamine synthetase genes of the pathogens examined. Both the glutamine synthetase and ${\beta}$-tubulin genes contained a complex set of polymorphisms. Based on these results, the pathogens isolated from organic cultivation orchards were found to have more diversity than the isolates obtained from the practical management orchards.

• Archives of Pharmacal Research
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• v.23 no.5
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• pp.488-494
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• 2000

Cerebellar granule and glial cells prepared from 7 day-old rat pups were used to investigate the effects of sub-acute nicotine exposure on the glutamatergic nervous system. These cells were exposed to nicotine in various concentrations for 2 to 10 days in situ. Nicotine-exposure did not result in any changes in cerebellar granule and glial cell viability at concentrations of up to 500 $\mu\textrm{M}$. In cerebellar granule cells, the basal extracellular levels of glutamate, aspartate and glycine were enhanced in the nicotine-exposed granule cells. In addition, the responses of N-methyl-D-aspartate (NMDA)-induced glutamate release were enhanced at low NMDA concentrations in the nicotine-exposed granule cells. However, this decreased at higher NMDA concentrations. The glutaminase activity was increased after nicotine exposure. In cerebellar glial cells, glutamate uptake in the nicotine-exposed glial cells were either increased at low nicotine exposure levels or decreased at higher levels. The inhibition of glutamate uptake by L-trans-pyrollidine-2,4-dicarboxylic acid (PDC) was lower in glial cells exposed to 50 $\mu\textrm{M}$ nicotine. Glutamine synthetase activity was lower in glial cells exposed to 100 or 500 $\mu\textrm{M}$ of nicotine. These results indicate that the properties of cerebellar granule and glial cells may alter after subacute nicotine exposure. Furthermore, they suggest that nicotine exposure during development may modulate glutamatergic nervous activity.

• KSBB Journal
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• v.5 no.2
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• pp.151-158
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• 1990

Investigations on the liability of nitrogen usuage by Nostoc muscorum that has nitrogen fixing ability, and cultured tobacco cells as they were associately cultured on nitrogen-free media and effects of polyamine on the associated culture condition were carried out. In addition, measurement on the activity of nitrate reductase, glutamine synthetase, glutamate dehydrogenase and glutamate synthase that take part in the metabolic pathway of nitrogen fixation product were performed. Among enzymes participating in the metabolic pathway of nitrogen fixation products, the activity of nitrogen reductase stimulated five times in associated culture, and that of glutamine synthetase of N. muscorum increased two times after heterocyst differentiated. Activity of glutamate dehydrogenase increased markedly when cultured tobacco cells were solely incubated on nitrogen-free media, but inhibited when cultured associately. And, glutamate synthase was showed the highest activity in 0.1 mM of spermine treated group.

• BMB Reports
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• v.33 no.6
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• pp.514-518
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• 2000

Two open reading frames of Haemophilus influenzae, HI0572 and HI0751, showing homology to a yeast thioredoxin peroxidase II (TPx II) and an E. coli thiol peroxidase $P_{20}$, respectively, were cloned and expressed in E. coli, and then the proteins were subsequently purified and characterized. HI0751 protein showed the thioredoxin (Trx)-dependent peroxidase activity, whereas HI0572 protein showed glutathione-dependent peroxidase. The HI0572 is the first peroxiredoxin with glutathione peroxidase activity rather than thioredoxin peroxidase. Purified HI0572 and HI0751 proteins protected specifically the inactivation of glutamine synthetase by metal catalyzed oxidation (MCO) systems composed of $Fe^{3+}$, $O_2$ and mercaptans such as dithiothreitol, ${\beta}-mercaptoethanol$ and glutathione (GSH). Unlike the HI0751 protein, the HI0572 protein was more effective in protecting glutamine synthetase from inactivation by the $GSH/Fe^{3+}/O_2$ system. It seems that these unique properties of the HI0572 protein are due to the structure containing a glutaredoxin domain at it's C-terminal in addition to a peroxiredoxin domain.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.57-61
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• 1998

Soybean and tobacco tissue cultured with modified MS media containing 4 different ratio(as N) of nitrate to ammonium combination which were 3:0, 2:1, 1:2 and 0:3. The highest callus growth in soybean were observed in the 2:1 medium. The medium containing nitrate only was detrimental to soybean callus growth. Tobacco callus grown with nitrate-only grew as well as those in the 2:1 and very slowly with ammonium-only. In tobacco callus, the total nitrogen in the callus increased with the increase of nitrogen concentration in the medium, but in soybean callus, the opposite result was noted. Nitrate reductase activity in tobacco callus was high when grown with nitrate-only but low with ammonium-only. In case of soybean callus, nitrate reductase activity was high in the 2:1 and remarkably low in nitrate-only medium. Both in soybean and tobacco callus, the activity of glutamine synthetase was high with nitrate-only, but low with ammonium.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.5
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• pp.515-521
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• 1997

Tobacco plant was grown for 40 days hydroponically in nutrient solutions composed of different forms of nitrogen, like NO$_3$$^{[-10]}$ -N, NH$_4$$^{＋}$-N, and a mixed formulation of NO$_3$$^{[-10]}$ -N and NH$_4$$^{＋}$-N. Uptake response, nitrate reductase, and glutamine synthetase activity at growth stage were investigated to understand the basic knowledge of nitrogen metabolism. The better growth of shoot and root was observed in the mixed nutrient solution than NO$_3$$^{[-10]}$ -N or NH$_4$$^{＋}$-N, alone. The plant growth in NH$_4$$^{＋}$-N nutrient solution was poor due to ammonium toxicity. The pH of nutrient solution containing NO$_3$$^{[-10]}$ -N increased up to 40 days after transplanting. But the pH of solution containing NO$_3$$^{[-10]}$ -N decreased drastically to 3.42 at 20 days after transplant. The pH in the mixed formulation dropped to pH 3.64 at 30 days after transplant and showed re-increase. It is assumed that nitrogen of NH$_4$$^{＋}$-N form was taken up preferentially at early stage and NO$_3$$^{[-10]}$ -N form was taken up preferentially at middle stage in the treatment with the mixed solution. The result indicates that the relative proportion of nitrogen forms affected the uptake patterns at each growth stages. The contents of chlorophyll and soluble protein were high with the mixed solution. Total nitrogen content was the highest in NH$_4$$^{＋}$-N solution and the content also increased by the application of the mixed type of nitrogen. The amount of nitrate in leaves was high in NO$_3$$^{[-10]}$ -N treatment and the amount of ammonium was high in NH$_4$$^{＋}$-N treatment. The activity of nitrate reductase or glutamine synthetase was highest in the leaves grown in mixed nutrient solution than in those with any other single of nitrogen form.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.210-210
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• 1996

Changs in the glutamatergic nervous system following AF64A injection into lateral ventricle were studied in rats. Rats were treated with the infusion of AF64A (3mM) into lateral ventricle At a week after the infusion of AF64A into lateral ventricle, rats were sacrified and each brain resions was dissected ; striatum, hippocampus and frontal cortex. At these resions, total glutamate and other amino acids levels. [$^3$H]Mk801 binding sites and glutamine synthetase activity were measured using HPLC-ECD, ligand binding assay and enzyme activity assay, respectively. The levels of total glutamate were decreased in striatum, hippocampus and frontal cortex Also, the levels of total glycine and taurine were decreased in all examined regions. Furthermore, the levels of total aspartate and GABA were decreased in both hippocampus and frontal cortex but these didn't alter in striatum. Additionally, the levels of total glutamine were decreased in both striatum and frontal cortex. The u\numbers of [$^3$H]MK801 binding sites were differently dffected in each brain resions ; the decrease in striatum, the increase in frontal cortex and no change in hippocampus Glutamine synthetase activity in striatum was significantly decreased. But, that in both hippocampus and frontal cortex didn't alter These results suggest that changes in the glutamatergic nervous system in three regions are induced by following AF64A injection into lateral ventricle in rats.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.296-300
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• 1990

For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.