• 약학회지
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• 제30권6호
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• pp.343-347
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• 1986

Korean ginseng was purified to obtain radioprotective protein fractions by buffer extraction, ammonium sulfate fractionation, CM-cellulose column chromatography, heat inactivation and Sephadex G-75 column chromatography. The final three fractions, GI, GII and GIII were subjected to Disc-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The molecular weights(M.W.) of native and denatured proteins were estimated by using regression line equations obtained from the mobilities of standard proteins. As the results, in Disc-PAGE, the GI fraction showed two protein bands with M.W. of above 213, 000 and 55, 000, GII showed one band with M.W. of 44, 000 and GIII, also one band with M.W. of 19, 000. In SDS-PAGE, GI fraction gave four subunit bands with M.W. of above 114, 000, 27, 000, 24, 000 and 19, 000, GII gave two bands with M.W. of 46, 000 and 22, 000, and GIII, one band of 19, 000.

• 미생물학회지
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• 제24권3호
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• pp.263-269
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• 1986

Membrane proteins of facultatively anaerobic Escherichia coli K12 which was logarithmically grown in aerobiosis and anaerobiosis were compared on 5 to 10% liner gradient gel electrophoresis (Na Dod $SO_4 -PAGE$). Membrane proteins were formed as different patterns between aerobiosis and anaerobiosis. Among them, 91Kdal protein (pbp1a) was not synthesized in aerobiosis and 60Kdal protein (fts cluster), in anaerobiosis. Thereby cells cultured aerobically were differenciated as diversiform cell shape, comparing cells cultured anaerobically and the latter were resistant to penicillin G. Thus it is believed that in facultative anaerobes atmospheric oxygen regulated the synthesis of membrane proteins and even the expression of equivalent genes, and moreover alleviated the resistance to an antibiotic penicillin.

• 약학회지
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• 제27권4호
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• pp.315-320
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• 1983

Ginseng proteins have been extracted and partially purified by the methods of ammonium sulfate fractionation, heat inactivation and Sephadex G-75 column chromatography. The final three fractions obtained (GI, GII and GIII) were subjected to paper chromatography and SDS-polyacrylamide gel electrophoresis. Molecular weights of polypeptide chains from each fraction were est-mated by the standard line obtained from the plot of electrophoretic mobilities against the logarithm of molecular weights of standard proteins. Results are as follows: 1) GI showed three protein spots and four polypeptides of which M.W. were 63,000, 60,000, 56,000, 51,000 and 34,200. 2) GII showed four protein spots and five polypeptides with their M.W. of 64,600, 56,000, 45,400, 35,800, end 25,200, 3) GIII showed three protein spots and four polypeptides with their M.W. of 66,000, 63,000, 56,000 and 22,000.

• KSBB Journal
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• 제8권3호
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• pp.287-294
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• 1993

사람의 뇨로부터 유용한 단백질을 동시에 분리하 기 위한 목적으로 효과적인 통합 분리정제 공정이 고안되었다. Ultrafiltration 방법을 이용한 농축과정 과 pH침전법을 전처리 단계로 사용하였고, 이어서 g gel filtration과 흡착, 이온교환, 친화, 그리고 역상 C column을 선택적으로 혼합한 chromatography를 수행하였다. 이 공정으로 정제한 urokinase, epider­m mal growth factor, albumin은 각각 SDS-poly­a acrylamide gel 전기이동상에서 단일의 단백질 띠로 이동하였고, 자신의 단백질 활성을 유지하고 있었다. 이들 세 가지 목적 단백질의 전체 수율은 각각 48 % %, 17%, 46%로 나타났다.

• Journal of Ginseng Research
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• 제13권2호
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• pp.254-259
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• 1989

A radioprotective ginseng protein fraction was obtained from Korean white ginseng powder by the following isolation and purification procedures: Tris-HCI buffer extraction, 70% ammonium sulfate fractionation, CM-rellulosr column chromatography, heat inactivation and Sephadex G-75 column chromatography. This fraction was further purified by Sepharose 4B and Sephadex G-150 column chromatographies. Three fractions obtained were subjected to Native-PAGE and SDS-PAGE using gradient gels and the silver staining method. Molecular weights of the native proteins and their subunits were estimated.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.553-562
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• 2004

To investigate protective activity in pepper plants, which were pre-inoculated with arbuscular mycorrhizal (AM) fungi Glomus intra radices (Gi), against pathogenic strain Phytophthora capsici (Pc), pathogenesis-related (PR) proteins and antioxidant enzymes were examined. The growth of root and shoot was the highest in peppers inoculated with G. intraradices, compared with non-inoculated control plants and those challenged by the pathogen with and without mycorrhizae after nine days of infection. Mycorrhizal colonization rate was reduced by about 10% in pathogen-challenged plants, but disease pressure was reduced. The activities of PR proteins, $\beta$-1- 3-glucanase and chitinase, were increased in Pc-treated plants compared to Gi+Pc-treated plants in leaves, but those in roots were suppressed. Superoxide dismutase activity and $H_2O_2${/TEX> content in Gi+Pc and Pc-treated plants were gradually increased in leaves. However, those in roots continuously increased up to 5 days, and then decreased dramatically. Peroxidase activity in leaves and roots increased after P. capsici infection both in plants inoculated with or without G. intraradices. These results suggest that AM fungi, G. intra radices, potentially act as one of the protective agents against plant pathogens. Changes of PR proteins and antioxidative enzymes in mycorrhizae-inoculated pepper appear to be regulated differently in leaves and roots by pathogen infection.

• KSBB Journal
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• 제15권2호
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• pp.174-180
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• 2000

본 연구에서는 다른 host cell 에 비하여 여러 가지 장점을 가지고 있는 Methylotrophic yeast 중 Pichia pastoris와 Hans-enula polymorpha의 fed batch 실험을 통하여 유전자 재조합 단백질 발현최적조건을 구하여 각 균주의 유전자 재조합 albumin 발현 최적화 연구를 수행하였다. 글리세롤이 두 균주의 promoter의 AOX 1과 MOX promoter repression에 미치는 영향을 확인한 바 H. polymorpha가 P. pastoris보다 promoter repression이 심함을 알 수 있었다. 두 균주의 promoter를 induction시키는 최적 메탄올 농도는 P. pastoris의 경우 메탄올 8g/L, H. polymorpha의 경우는 13 g/L임을 알 수 있었다. 또한 메탄올에 의한 induction시기는 두 균주 모두 O.D. 4 정도되는 exponential growth stage에서 메탄올을 첨가하는 경우가 초기 세포 성장단계에 메탄올을 첨가한 경우에 비해 약 20% 정도 높아짐을 확인하였다. 두 균주의 재조합 albumin 발현에 미치는 pH의 영향을 조사하였는데, p. pastoris의 경우 pH 5에서 가장 높은 albumin 생산성을 보여 약 300mg/L albumin을 발현하였고, H. polymorpha 의 경우 pH 5와 6에서 최대 약 180 mg/L의 albumin을 발현하였지만 pH 8에서는 이의 절반 수준에 그쳤다. 두 균주의 최적 fed-batch 방법을 확인하는 실험을 수행하였는데 P. pastoris의 경우의 최적 fed-batch 방법은 mixed feeding은 바람직하지 않고 글리세롤 배지를 공급하여 세포를 성장시킨 후 글리세롤 공급을 멈추고 바로 메탄올로 전환하는 방법을 효과적이며, H. polymorpha의 경우 비성장속도 제어를 통한 글리세롤 공급으로부터 메탄올 공급으로의 단계적 전환방법이 균주의 albumin 발현에 큰 영향을 준다는 것을 알 수 있었다. 이 방법을 통하여 두 균주의 고농도 배양 실험을 수행한 결과 P. pastoris의 경우는 O.D. 300에서 약 4.7g albumin/L를 발현하였다. 이와같은 결과를 바탕으로 산업체에서 methlotrophic yeast를 이용한 상업화를 계획함에 있어서 host로서의 균주를 선택할 수 있는 기본 자료를 제공함과 아울러 균주가 선택된 후에 그 균주를 이용한 재조합 단백질 최적화 방법을 제공하여 줄 것으로 생각된다.

• 한국임상수의학회지
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• 제21권1호
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• pp.20-22
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• 2004

개의 아토피성 피부염의 주된 세균성 감염은 Staphylococcus intermedius에 의해 발생한다. 본 연구의 목적은 아토피성 피부염을 가지고 있는 환견의 혈청과 정상견의 혈청을 이용하여 체액성 면역반응을 유발하는 주된 세균단백질을 규명하는데 있다. 건국대학교 수의과대학 부속동물병원에 내원한 아토피성 피부염 및 표재성 세균성 농피증을 앓고 있는 환견의 혈청 및 정상견의 혈청을 분리하여 본 실험에 사용하였으며 아토피성 피부염을 가지고 있는 환견에서 S. intermedius 균을 분리하였다. Brain heart infusion 액체배지 조건 및 $37^{\circ}C$ 배양조건에서 호기상태로 증균시켰으며 증균후 포도상구균을 PBS완충액에 부유시킨후 원심분리하고 최종적으로 lysostaphin을 이용하여 세균단백질을 분리하였다. 수거한 세균단백질은 SDS-PAGE을 이용하여 단백질을 전기영동하였으며 영동후 nitrocellulose membrane을 이용하여 단백질을 이동시켰다. Western blot은 anti-dog-IgG, 아토피성 피부염의 혈청 및 정상혈청을 이용하였다. 결론적으로 아토피성 피부염의 혈청을 이용해 확인한 S. intermedius의 주요 세균단백질은 18, 31, 75, 및 110 kDa이였다. 현재의 연구결과로 볼 때 아토피성 피부염에 감염된 대부분의 환견은 S. intermedius의 세균단백질에 대한 체액성 면역반응이 유발된다고 생각된다.

• Journal of Animal Science and Technology
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• 제56권2호
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• pp.6.1-6.4
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• 2014

The aims of study were to investigate the effects of intraperitoneal (i.p.) infusion of ghrelin on pancreatic ${\alpha}$-amylase outputs and the responses of pancreatic proteins to ghrelin that may relate to the pancreatic exocrine. Six male Sprague-Dawley rats (300 g) were randomly divided into two groups, a control group (C, n = 3) and a treatment group (T, $10.0{\mu}g/kg$ BW, n = 3). Blood samples were collected from rat caudal vein once time after one hour injection. The concentrations of plasma ghrelin, cholecystokinin (CCK) and alfa-amylase activity were evaluated by enzyme immunoassay (EIA) kit. Two-dimensional gel electrophoresis (2-DE) analysis was conducted to separate the proteins in pancreas tissue. Results showed that the i.p. infusion of ghrelin at doses of $10.0{\mu}g/kg$ body weight (BW) increased the plasma ghrelin concentrations (p = 0.07) and elevated the plasma CCK level significantly (p < 0.05). Although there was no statistically significant, the ${\alpha}$-amylase activity tended to increase. The proteomics analysis indicated that some pancreatic proteins with various functions were up- or down-regulated compared with control group. In conclusion, ghrelin may have role in the pancreatic exocrine, but the signaling pathway was still not clear. Therefore, much more functional studies focus on these found proteins are needed in the near future.

• 미생물학회지
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• 제21권2호
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• pp.61-70
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• 1983

Synchronized cat kidney cells chronically infected with feline leukemia virus (FeLV) were used to study virus production, the synthesis of group specific antigen (gag) and envelope (env) proteins, the expression of env protein on the cell surface during the cell cycle, and the stability of viral RNA. As detecting method, we developed the radioimmunoassay (RIA) system using beta-emission of $^{131}I$ and demonstrated the validity of this system by comparison with routine RIA system using gamma-emission of $^{125}I$. The produced virus was analysed by developed RIA interval was determined by measuring reverse transcriptase activity. The results show that infected cells produce the complete virus particle containing products of gag, env and pol genes of FeLV, and maximum virus production occurs during mitosis of synchronized cells. Labeling of the cell surface of synchronized cells with $^{131}I$ shows that the amount of $gp70^{env}$ on the cell surface parallels cellular gorwth. Therefore, the cell cycle-dependent release of virus is not petition RIA of synchronized cells with $^{131}I$ labeled viral proteins synthesis during the cell cycle. The rate of synthesis of gag protein shows three peaks, corresponding to the $G_1,\;late\;S\;and\;late\;G_2$ phases of cell cycle. But the rate of synthesis of env protein dose not change, suggesting that in these cells the synthesis of these two gene products in controlled seperately. In Actionomycin D treated cells, the synthesis of viral proteins decreased sharply from 8 hours after treatment, and the late S and $G_2$ peaks of gag protein synthesis were disappeared. This shows the stability of viral RNA for about 6 hours in the absence of continuing viral RNA synthesis.