• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.141.2-142
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• 2002

• KSBB Journal
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• 제32권2호
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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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• 제31권2호
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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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• 제23권5호
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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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• 제5권2호
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• pp.151-158
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• 1990

부질소 1-B5 배지에서 단독 배양한 담배 배양세표의 nitrate reductase 활성은 1-B5 배지에서 배양한 담배 배양세포에 비해 50% 이상 감소되었으나 $10^{-4}$ sper-mine 처리구에서는 그 활성이 가장 증가되었으며, N.muscorum파 혼합 배양시 그 활성이 현저히 증가한 반면 polyamine은 거의 영향을 미치지 않았다. Glutamate dehydrogenase 는 혼합 배양시 담배 배양세포플 단독 배양하였플 때보다 약 4배 감소 되었으며, glutamate synthas$\xi$의 활성은 $10^{-4}M$ spermlne 처리구에서 혼합 배양 하였을 때 그 활성이 가장 높았다.

• BMB Reports
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• 제33권6호
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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.

• 식물조직배양학회지
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• 제25권1호
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• pp.57-61
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• 1998

본 실험은 질소원처리가 식물의 생육 및 질소대사에 미치는 영향을 규명하고자 조직배양을 실시하였으며 공시재료로는 담배 'NC 2326'와 대두 '단엽콩'을 사용하였다. 배지내 질소원의 종류와 그 비율을 달리했을 때 callus의 생육, 전 질소, 단백질 함량 그리고 nitrate reductase와 glutamine synthase의 활성 변화를 조사하였다. 단엽콩의 조직배양시 callus증식은 2:1처리구와 암모늄태 질소만 공급한 배지에서 높았던 반면 질산태만 공급한 경우 쉐는 callus증식이 저조하였다. 한편 담배는 질산태와 암모늄태 질소를 2:1로 혼용한 구가 callus증식이 좋았고, 암모늄태 질소단용구에서 저조하였으며, 질산태 질소단용구의 callus생육이 양호하였다. Callus내 전질소함량은 담배의 경우 배지내 질산태 질소가 많을수록 증가되어 질산태 질소단용구에서 가장 많았으며 암모늄태 질소단용구에서 가장 적었다. 한편 단엽콩은 이와 반대의 경향으로 암모늄태 질소단용구에서 가장 많았으며 질산태 질소단용구에서 가장 적었다. Callus의 nitrate reductase의 활성을 보면 담배는 질산태 질소단용구에서 활성이 높았으며 암모늄태 질소단용구에서 활성이 낮은 반면 단엽콩은 2:1 처리구에서 활성이 높았고 생육이 부진했던 질산태 질소단용구에서 현저하게 낮았다. Glutamine synthetase의 활성은 단엽콩, 담배 모두 질산태 질소단용구에서 활성이 높았으며 암모늄태 질소단용구에서 활성이 낮았다.

• 한국작물학회지
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• 제42권5호
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• pp.515-521
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• 1997

질소의 형태를 질산태(NO$_3$$^{-}$-N), 암모니아태(NH$_4$$^{＋}$-N) 그리고 질산태와 암모니아태(NO$_3$$^{-}$-N ＋ NH$_4$$^{＋}$-N)를 혼합한 양액 하에서 수경재배하여 담배의 생육단계별 질소이용에 따른 흡수 양상과 생육 및 nitrate reductase, glutamine synthetase 효소활성을 평가하여 질소 시비 유형에 따른 체내 질소대사의 기초자료를 제공하고져 축행한 결과는 다음과 같다. 질소의 형태별 생육은 NO$_3$$^{-}$-N, NH$_4$$^{＋}$-N 단독 처리구보다 혼합처리구에서 지상 및 지하부의 생육이 좋았고, NH$_4$$^{＋}$-N 처리구에서는 암모늄 독성에 의해 생육이 극히 불량하였다. 생육단계별 양액의 pH 변화는 NO$_3$$^{-}$-N 처리구에서 이식 초기부터 이식 후 40일까지 증가하였고 NH$_4$$^{＋}$-N 처리구에서는 이식 후 20일에 pH 3.42, 혼합처리구에서는 이식 후 30일에 pH 3.64까지 떨어졌다가 증가하였다. 혼합처리구의 질소흡수반응은 생육초기에서는 NH$_4$$^{＋}$-N의 흡수를 우선하고 생육중기부터 NO$_3$$^{-}$-N 흡수가 증가하여 흡수 양상을 달리하였다. 생육시기에 따라 흡수형태도 NO$_3$$^{-}$-N와 NH$_4$$^{＋}$-N 사이의 상대적인 비율에 의해 크게 영향을 미쳤다. 엽록소와 가용성 단백질은 혼합처리구에서 전질소는 NH$_4$$^{＋}$-N 처리구에서 높았고, 특히 NH$_4$$^{＋}$-N 질소를 시비하였을 때는 질소함량이 증가하였다. 질산함량이 증가하였다. 질산함량은 NO$_3$$^{-}$-N 처리구에서 암모니아함량은 NH$_4$$^{＋}$-N 처리구에서 높았고, 산소 활성은 혼합처리구에서 nitrate reductase와 glutamine synthetase 활성이 높았다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1996년도 춘계학술대회
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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.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.296-300
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• 1990

자외선 조사와 NTG를 처리하여 Brevibacterium flavum 10AHR(arg his $Rif^r$)과 Corynebacterium glutamicum 11TS(trp $Sm^r$의 돌연변이주를 분리하였다. B.flavum 10AHR과 C.glutamicum 11TS를 300$\mu g$/ml의 lysozyme으로 18시간 처리하여 원형질체를 형성하고, 융합시 30의 PEG 6,000으로 처리하였을 때 가장 높은$3.7\times 10^{-6}$의 융합빈도를 나타내었다.