• 한국초지조사료학회지
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• 제29권2호
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• pp.103-110
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• 2009

유채(Brassica napus L.)에서 황 결핍이 $NO_3^-$ 흡수와 동화에 대한 영향을 알아보고자 $SO_4^{2-}$ 농도를 세가지 수준(1 mM $SO_4^{2-}$, 대조구; 0.1mM $SO_4^{2-}$, 결핍; 0 mM $SO_4^{2-}$, 무공급)으로 25시간 처리한 후 $NO_3^-$ 흡수량, 식물조직내의 nitrate reductase (NR) 및 glutamine syntheetase (GS) 활성을 분석하였다. 25시간 처리과정에서 황결핍 조건하에서의 $NO_3^-$의 흡수는 대조구와 큰 차이를 나타내지 않은 반면, 황 무공급구에서는 $NO_3^-$의 흡수는 유의적으로 감소하였다. NR 활성은 잎과 뿌리 조직에서 공히 $SO_4^{2-}$ 농도가 감소됨에 따라 감소하였다. 특히 어린잎과 중간 잎의 황 결핍 처리구에서 각각 35% 및 황 무공급구에서 70%의 뚜렸한 감소를 보였다. 뿌리에서는 오직 황 무공급구에서 유의적인 감소 (-29%)를 보였다. $SO_4^{2-}$ 농도에 따른 잎 조직에서 GS 활성은 잎의 성숙 정도에 따라 차이를 보였으며, 전반적으로 황 결핍과 황 무공급구에서 감소하는 경향을 보였다. 뿌리조직에서 GS 활성은 황 무공급구에서만 유의적인 감소를 보였다. 이러한 결과들은 황 공급이 제한을 받는 조건에서 질산염의 흡수가 감소되며, 식물조직체내의 질산염환원과 아미노산 합성관련 효소의 활성이 제한을 받았다는 것을 보여준다.

• BMB Reports
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• 제29권3호
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• pp.236-240
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• 1996

A 23-kDa molecular mass of antioxidant protein was purified from human liver. This protein exhibited the preventive effect against the inactivation of glutamine synthetase by a metal-catalyzed oxidation system. This antioxidant activity was supported by a thiol-reducing equivalent such as dithiothreitol in a similar manner to that of the 25-kDa thiol-specific antioxidant protein (TSA) from human red blood cells (HR). However, a thioredoxin-linked peroxidase activity of thiol-specific antioxidant protein of human liver (HLTSA) (0.91 ${\mu}mol/min/nmol$ of HLTSA) was much lower than that of thiol-specific antioxidant protein of human red blood cells (HRTSA) (16.4 ${\mu}mol/min/nmol$ of HRTSA). This HLTSA is also immnologically distinct from HRTSA Amino acid sequences of the three tryptic peptides (P1, P2, P3) of HLTSA were found to be completely homologous to segments of the known Mer5-like protein, which belongs to the known TSA family.

• 미생물학회지
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• 제26권2호
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• pp.93-100
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• 1988

In order to understand the regulation of glutamate dehydrogenase(GDH) synthesis in Brevibacterium flavum, we have isolated a mutant lacking NADP-linked GDH activity by ethlmethane sulfonate treatment. The $gdh^-$ mutant was grown on the minimal plate with 1mM ammonium chloride and not that with 300mM ammonium chloride. The cell-free extracts from $gdh^-$ mutant and prototroph were also examined with glutamine synthetase(GS) and glutamate synthase (GOGAT) production by niteogen sources. The growth of $gdh^-$ mutant in presence of 20mM ammonium chloride means that GOGAT synthesis is sufficient to allow growth in this condition. GS production of $gdh^-$ mutant as well as parental strain was induced by 1mM urea and ammonium tartrate, but it was repressed by higher concentration of ammonia, and also induced by 20mM to 50mM glutamate as a substrate. It was special attention that GOGAT synthesis from $gdh^-$ strain was more repressed by higher concentration of ammonia than prototroph as described in E. coli system.

• 한국환경성돌연변이발암원학회지
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• 제8권1호
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• pp.22-34
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• 1988

The biochemical changes of the hepatic tissues, induced by the carcinogen treatment such as diethylnitrosamine and acetamidofluorene in combination with the partial hepatectomy after Solt and Farber, were determined for the characterization of the induction of the proliferative capacity and the environmental adaptability of the carcinogenic tissues during the malignant transformation process. For the study of the proliferative capacity of the tissues, the activities of the enzymes, related with the nitrogen trapping mechanism, such as glutamine synthetase and gamma-glutamyltranspeptidase, were monitroed, while the cintents of cytochrome P450's and their isozymic patterns as well as the activities of the glutathione S-transferase were determined in the function of time after the hepatocarcinogenic stimuli.

• 대한간호학회지
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• 제28권4호
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• pp.893-907
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• 1998

The purpose of this study was to determine the effect of regular exercise during dexamethasone injection on the body weight, weight of hindlimb muslces, myofibrillar protein content and glutamine synthetase activity. 180-200g female Wistar rats were divided into four groups : control, exercise, dexamethasone injection (dexa), and exercise during dexamethasone injection(D+E) group. The dexa group received daily subcutaneous injection of dexamethasone at a dose of 4mg/kg body weight for 7 days. The exercise group ran on a treadmill for 60min/day(20minutes every 4 hours) at 10m/min and a 10$^{\circ}$grade. The control group received daily subcutaneous injection of normal saline at a dose of 4mg/kg body weight for 7 days. The D+E group ran on a treadmill for 60min/day(20minutes every 4 hours) at 10m/min and a 10$^{\circ}$ grade during dexamethasone injection. Body weight of the control group increased significantly from days of experiment, that of the dexa group decreased significantly from day 4 of the experiment resulting in a 82.4% decrease compared to the first day of the experiment. Body weight of the D+E group decreased significantly from day 5 of experiment resulting in a 81.77% decrease comprared to the first day of the experiment. Body weights, muscle weight and myofibrillar protein content of the plantaris and gastrocnemius decreased significantly and muscle weight of the soleus tended to decrease with dexamethasone injection. Glutamine synthetase activity of the hindlimb muscles increased significantly with the dexamethasone injection. The relative weight of the soleus was comparable to the control group and that of plantaris decreased significantly and that of gastrocnemius tended to decrease compared to that of the control in the dexa group. Body weight and muscle weight of the plantaris and gastrocnemius of the excrcise group were comparable to the control group, and the muscle weight of soleus showed a tendencey to increase. The relative weight of the soleus increased significantly and that of the plantaris and gastrocnemius were comparable to the control in the exercise group. Myofibrillar protein content of the soleus and plantaris increased significantly and there was no change of GS activity of the hindlimb muscles compared to the control in the exercise group. Body weight of the D+E group was comparable to the dexa group, muscle weight of the plantaris increased significantly and that of the soleus and gastrocnemius showed a tendency to increase. The relative weight of the hindlimb muscles increased significantly. Myofibrillar protein content of the soleus and plantaris increased significantly and that of the gastrocnemius tended to increase compared to the dexa group. Body weight and muscle weight of the plantaris and gastrocnemius of the D+E group did not recover to that of the control group. Muscle weight of the soleus recovered to that of the control group. The relative weight and of myofibrillar protein content of the hindlimb muscles recovered to that of the control group. From these results, it is suggested that regular exercise during dexamethasone injection might attenuate the muscle atrophy of the hindlimb muscles.

• 식물조직배양학회지
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• 제21권2호
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• pp.99-103
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• 1994

비선택성 제초제인 bialaphos는 고등식물에 있어서 glutamine 합성을 억제하여 식물체를 고사 시키는 능력을 갖고 있다. 본 연구에서 acetylteansferase에 의해 encoding된 bialaphos 저항성 유전자(bar gene)는 세균(Pseudomonas sryngae pv tabaci)의 genomic DNA로부터 cloning된 것을 사용하였다. 수분시킨 담배의 화계에 일정한 시간별로 bar 유전자를 처리한 결과 수분 후 30-40시간 사이의 처리구 에서 형질전환 식물체가 가장 많이 얻어 졌다. 그러한 형질전환 식물체의 kanamycin과 bialaphos 저항성 형질은 자식후대(T$_1$, T$_2$)에 있어서도 우성형질로 유전되었으나 wild type의 담배는 상기의 약제를 처리 하였을때 전부 고사하였다. 그리고, T$_1$세대의 형질전환 식물체로부터 전 염색체 DNA를 추출하여 Southern 분석한 결과 bar 유전자가 식물의 염색체상에 안정하게 존재하는 것을 확인하였다. 이상의 결과로부터 담배의 수분, 수정 시기에 외부유전자인 bar를 화주에 처리함으로써 bialaphos 저항성 식물을 만들어낼 수 있었다.

• 자연과학논문집
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• 제14권2호
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• pp.67-82
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• 2004

고염에서 자라는 원시 박테리아인 Halococcus agglomeratus에서 Thiol-specific antioxidant 활성을 보이는 분자량이 22-kDa인 향산화 단백질을 순수 분리 정제하여 향산화 활성의 특성을 조사하였다. 그 결과 진핵 세포의 Thiol-specific antioxidant protein (TSA of TPx)과 유사한 활성을 갖는 것을 확인 할 수 있었다. 정제된 Thiol-specific antioxidant protein 은 환원제로 thiol 성분을 갖는 비효소적 금속 촉매 산화계( $Fe^{3+}$, $O^2$, DTT 또는 2-mercatoethanol : thiol- MCO system)에 의하여 Glutamine Synthetase (GS)의 불활성화를 방어하고 Ascorbate 같은 nonthiol 성환원제를 갖는 금속 촉매 산화계 ( $Fe^{3+}$, $O^2$, Ascorbatenol: nonthiol- MCO system)에 의해서는 GS의 불활성화를 방어하지 못하였다. 이것은 환원형 thiol성분이 항산화 단백질의 항산화 활성에 전자 공여체로 요구되어 지기 때문이라고 판단된다. 이 단백질은 다른 TPx와는 다르게 100%의 활성을 나타내려면 NaCl의 농도가 500mM이상이 되어야 한다. 이상의 결과는 원시 박테리아에도 TPx가 존재하여 활성 산소종을 제거하는 생화학적 역할을 수행함을 시사하고 있다.

• Journal of Microbiology
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• 제44권3호
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• pp.301-310
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• 2006

Two pathways of ammonium assimilation and glutamate biosynthesis have been identified in microorganisms. One pathway involves the NADP-linked glutamate dehydrogenase, which catalyzes the amination of 2-oxoglutarate to form glutamate. An alternative pathway involves the combined activities of glutamine synthetase, which aminates glutamate to form glutamine, and glutamate synthase, which transfers the amide group of glutamine to 2-oxoglutarate to yield two molecules of glutamate. We have cloned the large subunit of the glutamate synthase (GOGAT) from Salmonella typhimurium by screening the expression of GOGAT and complementing the gene in E. coli GOGAT large subunit-deficient mutants. Three positive clones (named pUC19C12, pUC19C13 and pUC19C15) contained identical Sau3AI fragments, as determined by restriction mapping and Southern hybridization, and expressed GOGAT efficiently and constitutively using its own promoter in the heterologous host. The coding region expressed in Escherichia coli was about 170 kDa on SDS-PAGE. This gene spans 4,732 bases, contains an open reading frame of 4,458 nucleotides, and encodes a mature protein of 1,486 amino acid residues (Mr =166,208). The EMN-binding domain of GOGAT contains 12 glycine residues, and the 3Fe-4S cluster has 3 cysteine residues. The comparison of the translated amino acid sequence of the Salmonella GOGAT with sequences from other bacteria such as Escherichia coli, Salmonella enterica, Shigella flexneri, Yersinia pestis, Vibrio vulnificus and Pseudomonas aeruginosa shows sequence identity between 87 and 95%.

• KSBB Journal
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• 제13권3호
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• pp.295-302
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• 1998

Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.

• Journal of Plant Biotechnology
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• 제42권4호
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• pp.350-355
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• 2015

Clubroot disease is one of the most wide-spread and devastating diseases in the cultivation of Chinese cabbage. To develop a protein marker for resistance to clubroot disease in Chinese cabbage, a comparative proteome analysis was performed between a sensitive line, 94SK, and a resistant line, CR Shinki DH. Three proteins of two fold or higher accumulation that are specific to each line were found 3 days after innoculation of the Plasmodiphora brassicae. They are glutamine synthetase, malate dehydrogenase/oxidoreductase and fructose-bisphosphate aldolase in the 94SK and actin, phosphoglycerate kinase, and Cu/Zn superoxide dismutase in the CR Shinki line. From the comparison of the synthesized proteins in the 94SK and the CR Shinki, CR Shinki was found to produce more ATP-binding protein for the ABC transporter while 94SK showed a higher level of pathogenesis-related protein 1 production. All of these proteomic variations may lead to the development of molecular markers to accelerate the breeding process.