• Parasites, Hosts and Diseases
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• 제30권2호
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• pp.133-140
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• 1992

T. sergenti merozoite 수용성 항원을 T. sergenti 감염적혈구로부터 분리하고자 저삼투압액으로 용혈, 조직 분쇄기로 분쇄한 후에 고속 원심분리하여 수용성 항원을 얻었으며, SDS-PAGE와 Western blot의 방법으로 29, 34, 35 그리고 105 kD가 함유된 항원을 본 예방접종실험의 항원성 polypeptide로 정하였다. 본 수용성 항원(0.5 mg/ml)을 준비, Freund's adjunant를 이용하여 한우(5개월령)에 경피 접종하였으며, 다시 4주 후에 추가접종하였다. 추가접종 9주 후에 예방 접종군과 대조군에 동종의 냉동충주(5.6$\times$106RBC/dose, 40% 기생률)을 접종시킨 후에 적혈구용적비, 총적혈구수, 기생률, western biot에 의한 특이항체 그리고 간접형광항체(IFA) 등을 관찰하였던 바, 예방접종 후 18주(충 접종 6주 후)에 있어서 예방접종군의 IFA는 10,240이었으나, 대조군은 1,280이었다. 예방접종군에 있어서의 충접종 전후에 있어서의 총적혈구소와 적혈구웅적비는 유의적 차이 (p<0.05)를 나타내지 않았지만, 대조군에 있어서는 적혈구용적비와 총적혈구수에서 있어서 빈혈 소견을 관찰하였다 (p<0.05). 예방접종군의 충전종 후에 있어서의 western blot 반응에서는 29, 34, 35 그리고 105 kD polypeptide의 물질이 면역반응을 잘 나타내고 있어, 이들 polypeptide는 앞으로 vaccine 제조에 활용 가능성이 충분함을 예견할 수 있었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.482-493
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• 2005

Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.

• Applied Biological Chemistry
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• 제29권1호
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• pp.73-77
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• 1986

Auxin과 cytokinin 그리고 $GA_3$가 대두(Glycine max) 종자의 단백질 합성과 축적에 미치는 영향을 보기 위하여, 2,4-D와 BA 그리고 $GA_3$을 각각 $10^{-6},\;10^{-5},\;10^{-4}M$의 수용액으로 만들어 개화기에 식물체 전체에 살포 처리하고 개화 후 33일의 미숙종자와 77일의 완숙종자에 대한 단백질 전기영동 양상을 조사하였다. $GA_3$는 단백질 합성과 축적에 영향을 주지 않았으나 2,4-D와 BA는 일정농도에서 특정 단백질들의 축적에 영향을 주었다. 7S의 ${\alpha}$와 ${\alpha}'$ subunit, 11S의 acidic subunit 들이 2,4-D 혹은 BA처리에 의하여 나타나지 않았으며, 이 subunit들이 성숙종자에 없으나 미숙종자에는 있는 점으로 보아, 2,4-D나 BA는 이들의 함성을 저해하는 것이 아니고 축적과정의 어느 단계에서 이 subunit들의 분해 혹은 전환을 촉진하는 것으로 생각되었다. 2,4-D와 BA에 의해 위의 단백질이 성숙종자에 축적되지 않는 것은 어떤 단백질분해효소(metalloendopeptidase?)의 작용과 관련이 있는 것으로 보이며, 2,4-D나 BA는 성숙후기에 이 효소의 gene expression을 촉진하거나 활성을 증진시키는 것으로 생각되었다.

• 한국식품과학회지
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• 제29권3호
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• pp.417-426
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• 1997

미강을 혈중 콜레스테롤 저하 효과가 있는 기능성 식품소재의 원료로 활용하는 데 필요한 기초 자료를 얻기 위하여 미강 단백질의 가수 분해물을 제조하여 담즙산 결합 획분을 분리하고 일부 물리화학적 특성을 조사하였다. 미강 단백질을 탈지 미강으로부터 알칼리 추출과 등전점 침전 방법을 이용하여 제조하였다. 미강 단백질을 기질로 하여 pH-drop method로 측정한 효소의 상대적 활성과 가수분해시 가수분해율의 변화를 비교 평가하여 기질의 가수분해에 적합한 효소를 선택하였다. Esperase에 의한 가수분해물을 한외여과(MWCO : 10 kDa)하여 두 부분으로 나누었다. 각 분획물을 cholic acid를 공유결합 시킨 ${\omega}-aminohexyl$ Sepharose 4B column에 걸어, 소수성 상호작용에 의해 cholic acid와 결합하는 폴리펩티드 및 펩티드를 deoxycholate 완충용액으로 용출시켜 분리하였다. 겔투과 크로마토그래피(Sephadex G-50)를 이용하여 한외여과 여과백의 담즙산 결합물에 대해 분자량 분포를 측정한 결과, 대부분 $2\;kDa{\sim}10\;kDa$에 걸쳐 넓게 분포하였고 일부는 $0.2\;kDa{\sim}0.6\;kDa$사이에 존재하였다. 한외여과 잔류액의 담즙산 결합물에 대해서는 preparative reverse phase HPLC를 실시하여 미강 단백질 가수분해물에서 3개(R-1, 2, 3)의 Peak를 분리하였다. 각 peak의 총 아미노산과 유리 아미노산 조성을 분석하여 단백질, 폴리펩티드 및 펩티드 부분의 아미노산 조성을 조사하였다. 그 결과 미강 단백질 가수분해물에서 얻은 peak의 경우 proline 함량이 미강 단백질의 4배에 달했고, 평균 소수도가 높은 peak일수록 유리 아미노산이 함량이 높았으며 평균 소수도는 미강 단백질보다 다소 높은 것으로 나타났다.

• Applied Biological Chemistry
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• 제28권3호
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• pp.187-195
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• 1985

당근에서 cytokinin, auxin 또는 GA에 의해 조절되는 polypeptide 및 단백질들을 동정하기 위하여, kinetin, BA, IAA, NAA 혹은 $GA_3$가 각각 $10^{-4},\;10^{-5},\;10^{-6}M$인 배지에서 suspension culture 한 당근 callus들의 추출물의 전기영동양상을 조사하였다. 생장조절제를 처리하지 않은 callus에서 15개 polypeptide band가 관찰되었는데, 이들의 분자량은 각각 $18._4,\;20._2,\;24._0,\;34._9,\;35._7,\;37._4,\;40._3,\;42._2,\;44._1,\;44._4,\;49._3,\;55._0,\;56._6,\;58._1$ 그리고 $59._9KD$였다. Kinetin, BA, IAA, NAA 그리고 $GA_3$에 의하여 합성이 촉진되는 것으로 보이는 polypeptide band는 각각 2개, 6개, 1개, 2개 그리고 4개였고, 합성이 억제되는 것으로 보이는 것은 각각 5개, 3개, 4개, 3개, 그리고 2개였다. 분자량 $40._3KD$ 및 $42._2KD$의 polypeptide들은 cytokinin류 생장조절제에 의하여 합성이 촉진되는 것으로 나타났고, $44._1KD$의 것은 auxin류에 의하여 합성이 촉진되며 $37._4,\;44._4,\;56._6KD$의 것은 auxin류에 의하여 합성이 억제되는 것으로 나다났다. Kinetin, BA, IAA, NAA 및 $GA_3$처리에서 상대 이동도가 0.56, 0.84, 0.92인 단백질 band들이 증가된 것이 관찰되었으나, 각 생장조절제들에 특유한 단백질은 동정할 수 없었다.

• Fisheries and Aquatic Sciences
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• 제2권1호
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• pp.85-92
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• 1999

We examined the expression patterns of heat shock proteins in primary cultured hepatocytes from flounder (Paralichthys olivaceus) exposed to heat shock. The expression of hsp90, hsp70, hsp40, hsp30, and hsp27 was induced and major polypeptides were hsp70, hsp30 and hsp27. Northern blot analysis showed that expression of hsp70 was inhibited by transcriptional inhibitor actinomycin D, suggesting that expression of hsp70 gene is regulated at the transcriptional level. Prolonged exposure of cells to an elevated incubation temperature $(30^{\circ}C)$ induced the transient synthesis of hsp90, hsp70, hsp40, and hsp30 whereas maintenance of cells at a slightly higher incubation temperature $(32^{\circ}C)$ induced the continuous syntheses of these hsps. When cells were incubated at a higher temperatures $(35^{\circ}C\;or\;37^{\circ}C)$, the synthesis of hsps was almost similar to that of hsps in cells exposed to 32't except for the induction of hsp27 synthesis. These results that temperature and incubation time for optimum expression of each hsp during prolonged heat shock are different.

• Journal of Microbiology
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• 제41권3호
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• pp.259-261
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• 2003

The nsdD gene has been predicted to encode a GATA type transcription factor with the type IVb zinc finger DNA binding domain functions in activating sexual development of A. nidulans. In several allelic mutants of nsdD producing truncated NsdD polypeptides lacking the C-terminal zinc finger, the transcription level of nsdD gene was greatly increased. Also in an over-expressed mutant, the transcription under its own promoter was reduced. These results suggest that the expression of nsdD is negatively autoregulated. When the nsdD gene was over-expressed, cleistothecia were formed in excess amounts even in the presence of 0.6 M KC1 that inhibited sexual development of the wild type. Northern blot analysis revealed that the expression of nsdD was repressed by 0.6 M KC1. These results strongly suggest that the inhibition of sexual development by salts was carried out via the nsdD involved regulatory network.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.94-100
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• 2003

We studied the effect of cadmium on chlorophylls and rubisco activation in Canavalia ensiformis L. leaves. Chlorophyll levels were reduced by 5.0 ${\mu}$M Cd. Rubisco activity at 5.0 ${\mu}$M Cd was significantly smaller than that at no treatment. Rubisco Content showed patterns of change similar to rubisco activity. These data suggest that rubisco activity was associated with an amount of rubisco protein, and that the activation and induction of rubisco is inhibited by Cd. The degree of intensity of 50 and 14.5 kD polypeptides identified as the large and small subunit of rubisco by SDS-PAGE analysis at 5.0 ${\mu}$M Cd was significantly lower than that at control, indicating Cd had a e f-fect on both subunits. Under the assumption that effects of Cd on rubisco may be r elated to rubisco activase, in addition to, its activity and content we re determined . The rubisco activase activity at 5.0 ${\mu}$M Cd was more decreased than the control. A similar change pattern was also observed in content of rubisco activase. Remarkable differences in the intensitiy of both the 45 kD and 41 kD band were found between at control and Cd-treatment. These results suggest that the change in the levels of rubisco activase leads to a subsequent alter action of rubisco levels.

• Journal of Photoscience
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• 제9권2호
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• pp.55-58
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• 2002

The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.

• BMB Reports
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• 제42권9호
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• pp.541-551
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• 2009

Amyloidogenesis defines a condition in which a soluble and innocuous protein turns to insoluble protein aggregates known as amyloid fibrils. This protein suprastructure derived via chemically specific molecular self-assembly process has been commonly observed in various neurodegenerative disorders such as Alzheimer's, Parkinson's, and Prion diseases. Although the major culprit for the cellular degeneration in the diseases remains unsettled, amyloidogenesis is considered to be etiologically involved. Recent recognition of fibrillar polymorphism observed mostly from in vitro amyloidogeneses may indicate that multiple mechanisms for the amyloid fibril formation would be operated. Nucleation-dependent fibrillation is the prevalent model for assessing the self-assembly process. Following thermodynamically unfavorable seed formation, monomeric polypeptides bind to the seeds by exerting structural adjustments to the template, which leads to accelerated amyloid fibril formation. In this review, we propose another in vitro model of amyloidogenesis named double-concerted fibrillation. Here, two consecutive assembly processes of monomers and subsequent oligomeric species are responsible for the amyloid fibril formation of $\alpha$-synuclein, a pathological component of Parkinson's disease, following structural rearrangement within the oligomers which then act as a growing unit for the fibrillation.