• 대한약리학회지
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• 제29권2호
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• pp.275-282
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• 1993

Microsomal epoxide hydrolase (mEH)은 epoxide형 중간대사물을 해독화하는 효소이다. 본 실험실에서는 thiazole 또는 pyrazine을 rat에 투여할 때 mEH mRNA수준이 증가되고 mEH가 유도증가한다는 것을 밝힌바 있다(Carcinogenesis, Kim et al, 1993). 본 연구에서는 Thiazole처리를 한 rat의 간 microsome 분획으로 부터 DEAE-cellulose column chromatography를 이용하여 mEH를 순수분리하였고, 이를 SDS-PAGE분석 및 N 말단 amino acid 서열분석으로 확인하였다. Pyrazine처리를 한 rat의 간 microsome분획에서는 mEH와 더불어 이와 관련된 43 kDa 단백질이 함께 정제되었다. 정제된 thiazole 유도성 mEH를 토끼에 주사하여 항체를 생산하였고, 이 항체를 이용한 immunoblot 분석을 하였을 때 간 microsome 분획의 mEH가 thiazole투여군에서는 대조군에 비하여 10배, pyrazine 투여군에서 7배 증가하였다. Pyrazine처치한 rat의 간 microsome 분획에서는 mEH 관련성 43 kDa 단백질이 동시 유도증가하는 것을 면역화학적 반응으로도 확인하였다. 이때 Pyrazine으로 유도된 rat의 간 microsome 분획 또는 정제분획에 존재하는 43 kDa 단백질과 mEH의 비율은 1 : 15로 나타났다. 정제된 mEH와 43 kDa 단백질의 N 말단 amino acid 서열을 분석하였을때 43 kDa 단백질의 N 말단이 mEH와 동일하게 나타나 관련 단백질임을 확인하였다. 이러한 mEN 유도현상에 종차가 있는지를 알아보기 위하여 thiazole과 pyrazine을 각각 rabbit에 투여하였을 때 rabbit에서 는 mEH의 유도증가가 일어나지 않았으며, pyrazine 투여군에서 43 kDa 단백질의 증가는 관찰 되었다. 본 연구는 thiazole 또는 pyrazine 투여후 mEH 발현이 유도증가되며, pyrazine 투여 후에는 mEH 및 이와 관련된 43 kDa 단백질이 동시유도되고, 이러한 mEH 유도발현에 rat와 rabbit간에는 종차가 있음을 보여준다.

• Parasites, Hosts and Diseases
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• 제39권3호
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• pp.241-246
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• 2001

The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T: OFR without signal sequence and C-terminal hydrophobic sequence, S: N-terminal 2/3 parts of S, A: C-terminal 2/3 parts, P; N-terminal 1/3 part, X: middle 1/3 part Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-47 vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T,66 kDa for S, 52 kDa for A,53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with Al9 clone in SAGI of Toxoplasma gondii (Nam et at., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species .

• BMB Reports
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• 제30권1호
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• pp.21-25
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• 1997

The growth. freezing resistance and electrophoretic protein patterns of carrot callus cultures were investigated following treatment with NaCl for various' intervals at 20$^{\circ}C$. Following 7 day exposure to 250 mM NaCl. freezing tolerance increased, which was measured by 2.3.5-triphenyl tetrazolium chloride (TTC) assay and fresh weight was reduced compared to control cells. Changes of electrophoretic patterns of total and boiling stable proteins were investigated using one or two dimensional gel system. Several proteins with molecular weight of 43 and 21 kDa increased by NaCl treatment. The most prominent change was detected in 21 kDa protein. The steady state level of this protein increased in NaCl treated cells, but decreased in control cells. Twenty one kDa protein was detected only in the NaCl treated cell when boiling stable protein was analyzed. The isoelectric point of 21 kDa protein was identified as 5.7. The timing of increase of 21 kDa protein was correlated to freezing resistance which implied the role of this protein in the induction of freezing resistance of the cell.

• Parasites, Hosts and Diseases
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• 제35권2호
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• pp.87-94
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• 1997

우리 나라에서 가장 감염자가 많은 기생충인 간흡충의 현 감염에 특이한 항원 분획을 찾고자 면역이적법을 이용하여 감염자의 혈청을 검색하였다. 실험적으로 토끼에서 얻은 간흡충 성충의 조항원에 시스테인계 단백질분해효소 억제제인 I-64를 첨가하였을 때 큰 분자량의 분획을 가장 잘 보존하여 200-14 kDa의 범위에 20개 이상의 분획을 관찰하였다. 이 조항원을 이용하여 경북 상주군 주민의 혈청을 면역이적법으로 검사하였다 대변검사, 피내반응검사, 효소면역법검사(ELISA)를 이 용하여 검사한 73명 중 충란양성자 49명의 혈청 내에 특이 IgG와 IgE 항체가 생성되었고, IgM과 IgA 항체는 특이하게 반응하지 않았다. 여러 항원 분획 중에서 43, 34, 28-25 kDa 항원이 현 감 염에 특이한 분획이고. 항체와 반응한 94, 80, 72, 68, 64, 62, 52, 47, and 40 kDa 분획은 특이하지 않은 분획임을 확인하였다. 추후에 각 항원과 이에 대한 혈청반응에 대하여 특성을 더 연구 할 필요가 있으며, 특히 치료후 추적검사를 통하여 혈청내 항체의 소실을 구명하여야 할 것이다.

• 한국축산식품학회지
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• 제38권5호
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• pp.1101-1108
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• 2018

The aim of this study was to investigate the effects of dry-aging (DA) and the cooking process on the myofibril protein functionalities and in vitro digestibility of proteins in beef loin. Six sirloins from beef were dry-aged for 28 d, and the control group (n=6) was analyzed 2 d postmortem for this study. Dimensional changes (reduction of thickness and surface shrinkage) after cooking were significantly greater in the control group than the DA group, whereas the shear force of the DA group was significantly lower than that of the control. Effect of cooking on aggregation, hydrophobicity, and in vitro digestibility were significantly higher in the DA group than in the control. After cooking, the protein in DA sirloins was more oxidized than in the control samples. According to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis result, the low molecular weight bands (below 17 kDa) increased in the DA group, finding that the protein characteristics of dry-aged beef was affected by cooking.

• 대한수의학회지
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• 제46권1호
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• pp.13-19
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• 2006

Generally known psittacosis or ornithosis is a disease of birds caused by the bacterium Chlamydia psittaci. Humans are accidential hosts and are most commonly infected from avian sources. It raises hepatitis or neurosis. As major outer membrane protein (MOMP) of Chlamydia psittaci has been known to play a role in the avoidance of host immune defenses, research on developing a Chlamydia vaccine has focused on the MOMP. In this study, the gene encoding the major outer membrane protein (MOMP) of the Chlamydia psittaci strain 6BC was cloned and expressed in Escherichia coli strain M-15. The recombinant DNA was cloned by fusion prokaryotic expression vector pQE30-GFPII. Expression of the recombinant protein was performed in E. coli and was induced by IPTG. The size of expressed recombinant protein is 74.220 kDa (MOMP, 43.260 kDa; GFP expression region, 30 kDa; $6{\times}His$ tag, 960Da). This protein was purified by using his-tagging-inclusion body. Recombinant protein was reconfirmed through ELISA test and western blot with antibody against pQE30-GFPII. It will be useful antibody development.

• Parasites, Hosts and Diseases
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• 제42권4호
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• pp.169-174
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• 2004

This study was carried out to find out specific proteins from different organs of Clonorchis sinensis. Crude extract, organ-specific and excretory-secretory (ES) proteins were analyzed by immunoblot with infected human sera. The bands of 7- and 17 -kDa were main component of intestinal fluid and ES protein and commonly found in all organ-specific proteins. The 17-kDa protein was observed from ES antigen, intestinal fluid, eggs and sperms, 26- and 28-kDa proteins were from the uterus, vitellaria, and ovary, and 34-, 37-, 43- and 50-kDa proteins were mainly from the testis and sperms. Serum of mice immunized with sperms reacted to the 50-kDa protein by immunoblotting and immunohistochemical staining showed a positive reaction at the seminal receptacle and seminiferous tubule. The present results show that the 7-kDa protein is a common antigen of every part or organ of C. sinensis, but different organs express their specific antigenic protein bands.

• International Journal of Industrial Entomology and Biomaterials
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• 제18권2호
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• pp.139-142
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• 2009

The histological and transmission electron microscopic studies revealed the synthesis activity predominantly in the hypopharyngeal glands of the nurse bees. The biochemical analysis of both, the hypopharyngeal gland extract and royal jelly elucidated unequivocally the proteins and lipids as the major constituents. Further the SDS-PAGE of hypopharyngeal gland extract showed about 17 protein bands, perhaps 14.10, 20.00, 29.00 and 43.00 kDa predominantly while that of royal jelly revealed only two protein bands of 29.00 and 43.00 kDa molecular weight suggesting them as the major royal jelly proteins (MRJP). The lipid profile of royal jelly consists of triglycerides, cholesterol, HDL, LDL and VLDL.

• 아시안잔디학회지
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• 제4권1호
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• pp.12-23
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• 1990

The chlorophyll-protein complexes were separated from thylakoid membranes of Spinach oleracea and Zoysia japonica by two gel Systems of LiDodSO4-PAGE and LiDodSo4/Urea- PAGE under nondenaturing conditions. Seven chlorophyll~protein complexes of CPI*, CPI, CPII*. CP47, CP43, CP29 and CPII were fractionated from both S,oleracea and Zjaponica by LiDodSO4-PAGE. CPI, CP47 and CP43 contained more chlorophyll a than chlorophyll b. The patterns of their absorption spectra at room temperature were similliar to that of chlorophyll a, judging by their UV-spedtroscopy. On the other hand, CPII* and CPII contained approximately equim-olar quantities of chlorophyll a and b. Additional five chlorophyll-protein complexes not separated in the LiDodSO4-PAGE system were electrophoretically isolated from both S, oleracea and Zjaponica by LiDodSO4/Urca-PAGE. The chlorophyll-protein complex just above LRCII $\alpha$in the gel appears CCII-RC separeted recently. 23 kDa and 20 kDa cho-protein complexes is probably LHCIa and LHCIb as judged from their molecular weight. Two novel chlorophyll~protein complexes designated "CPI7" and "CPI6" were fractionate by this gel system. Their molecular weights respectively. Although the stoichiometry of their components and their roles in thylakoid membranes are not apparant, It is thought that they are another kinds of LHCI.other kinds of LHCI.

• BMB Reports
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• 제34권6호
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• pp.496-501
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• 2001

Lincomycin, an inhibitor of plastid protein synthesis, was found to block the synthesis of apoprotein P700 with a molecular mass of 72 kDa and the assembly of the Chl a-protein of PS I. Synthesis of the polypeptides of 48, 43.5, and 32 kDa of the PS II complex is also suppressed. This process is accompanied by the disappearance of the PS Two reaction center Chl a at 683 nm, and of the PS One reaction center Chl a at 690, 696, and 705 nm on the fourth derivative of the absorption spectra at 77K. Lincomycin does not affect the synthesis of LHC subunits. It increases the content of the two main Chl forms of LHC at 648 nm (Chl b) and 676 nm (Chl a). The low-temperature fluorescence ratio F736/F685 is also increased. However, the effect of cycloheximide (an inhibitor of cytoplasmic protein synthesis) leads to the reduction of polypeptides of the light-harvesting Chl a/b-protein complex in the range of 29.5-22 kDa. Under these conditions, the relative amount of Chl b and the F736/ F685 fluorescence ratio decrease significantly. This is obviously the result of blocking the LHC I and LHC II synthesis. At the same time rifampicin and actinomycin D (inhibitors which block transcription in chloroplast and nuclear genome, respectively) inessentially affect the characteristics of these complexes.