• 한국응용곤충학회지
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• 제34권2호
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• pp.100-105
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• 1995

정확한 in vitro 전사가 일어날 수 있는 진딧물의 세포추출액을 제조하였다. 전사를 직접 조절할 수 있는 단백질 인자를 규명하기 위하여 전사개시점과 그의 상류에 결합하는 DNA 결합단백질을 탐색했다. 전사개시점을 포함하는 단편 A(-194/23)에는 52kDa, 50kDa, 40kDa의 단백질들이 결합했으며 전사개시점 상류의 DNA 단편 B(-393/-263)에는 52kDa, 50kDa, 40kDa의 단백질들이 결합한 반면 DNA 단편 C(-263/-195)는 53kDa단백질만이 결합했다. 그리고 이들 DNA 결합단백질들의 DNA 결합 활성에는 양이온이 요구되었다.

• 한국미생물·생명공학회지
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• 제21권6호
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• pp.513-519
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• 1993

The alkaline protease in the polyhedra preparation of Spodoptera litura nuclear polyhedrosis virus was successfully inactivated by heating at 100C for 20 minutes. SDS-PAGE analysis indicated that heat inactivated polyhedra is composed of major proteins of 31kDa and presumptive its polymer protein of 62kDa. However, this polyhedra was converted into several smaller molecular weight proteins when treated with midgut juice, but not by treatment with heat-inactivated midgut juice.

• Archives of Pharmacal Research
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• 제18권5호
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• pp.301-307
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• 1995

In the particulate fraction obtained from PKC-down regulated K562 cells by treatment for 24 h with 200nM TPA, phosphorylation of two proteins with molecular weight, 100 kDa and 23 kDa (designated p100 and p23, respectvely) was depleted and addition of exogenous purified PKC to this fraction failed to testore their phosphorylation. However, in the soluble fraction, all of phosphoproteins abolished by long-term treatment with TPA were restored by exogenously added PKC. Phosphorylation of two proteins was increased by short-term tretment (20 min), and diminished with the persistant exposure to TPA as well as at a concentration as low as 1nM. When K562 cells were treated with 1 nM and 200 nM TPA for 24 h, phosphorylation of p100 was restored with or without exogenous PKC on 2-3day and 6day after removal of treated TPA, respectively. Two-dimensional electrophoresis of phosphoproteins revealed that phosphorylated p100 (pl=5.9) and p66 species were completely absent from the particulate fraction of K562 cells treated with 200nM TPA for 24 h. These results suggest that the interaction of sensitive endogenous substrates, p100 and p23 with the plasma membrane might be regulated by PKC-down regulation without displacement to the cytosol and the interaction of p100 with the membrane might be reveersible.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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• pp.432-437
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• 2002

Immunosuppressive potential of 65 kDa buffalo placental protein (bPP65) on B-cell proliferation in vitro and antibody response in vivo was evaluated. B-cell proliferation was estimated by measuring incorporation of tritiated thymidine in buffalo lymphocytes while primary antibody responses against phytohaemagglutinin (PHA) or keyhole limpet haemocyanin (KLH) were evaluated in mice. bPP65 suppressed proliferation of lipopolysaccharide (a B-cell specific mitogen)-stimulated buffalo lymphocytes in vitro indicating suppression of B-cells. This suppression was dose dependent over the protein concentration range $25-100 {\mu}g/ml$. Primary antibody responses in mice against PHA and KLH in presence of bPP65 were lower as compared to in its absence but these were not statistically significant. Amino acid composition data of bPP65 and BSA suggested that bPP65 is different from BSA.

• 대한수의학회지
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• 제36권2호
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• pp.453-461
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• 1996

Eighteen holstein-calves(4~5 months old) in a divided groups including the matched control were immunized with $100{\mu}g/dose$ of 34kDa, 45kDa polypeptide and T sergenti merozoite vaccine(protein content $100{\mu}g/dose$) respectively, previously mixed with aluminium hydroxide to elicit antibodies. All groups of calves were boosted with same dose and intervals. The animals were challenged by tick infestations in the endemic pasture of theileriosis from March to September 1994. The animals were monitored for the erythrocyte count, parasitemia, hematocrit and the specific antibody reactions elicited by immunization. The immunological responses demonstrated that vaccination with 34kDa polypeptide and T sergenti merozoite derived vaccine inhibited to produce the 75kDa band immunological responds even in the vaccinated calves after being challenged by tick infestations in the pasture. However, the specific antibody reactions were detected at the 32kDa band in the nonimmunized calves and T sergenti merozoite derived vaccine by the western blot. The 34kDa polypeptide vaccine and T sergenti merozoite derived vaccine were evaluated to be able to protect inducing anemia and to decrease parasitemias level. These vaccines have the efficacy of inhibition to produce a certain antigen corresponding 75kDa band antigen of parasite in the calves as challenged with tick infestations.

• 약학회지
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• 제44권1호
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• pp.47-51
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• 2000

To examine whether DGRF inhibits $cPLA_2$ activity in vitro, we purified a 100 kDa $cPLA_2$enzyme from porcine spleen and performed an inhibition study at two concentrations of 5.0 and 50.0 $\mu$M 1-stearoyl-2-[1-$^{l4C}$ ]arachidonoyl-sn -glycero-3-phosphocholine as a substrate to rule out an apparent inhibition due to "substrate depletion". Here we reported that DGRF inhibited $cPLA_2$activity with $ID_{50}$ of 3.2 $\mu$M and virtually complete inactivation of the enzyme occurred at 60 $\mu$M. Interaction experiment between enzyme protein and inhibitor by ultrafiltration method indicated that 1-desgalloylrugosin-F inactivates $cPLA_2$enzyme by an irreversible mechanism.

• 한국가축번식학회지
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• 제26권4호
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• pp.377-384
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• 2002

To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.805-810
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• 2000

An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

• BMB Reports
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• 제33권6호
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• pp.510-513
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• 2000

Boar sperminogen was purified from the acid extracts of the washed epididymal spermatozoa by gel filtration through a Sephadex G-100 column, followed by preparative SDS-PAGE. The 32 kDa sperminogen band was sliced out from the preparative SDS-PAGE and 32 kDa sperminogen was eluted from the gel matrix. The purified 32 kDa sperminogen was subjected to amino acid composition analysis. The amino acid composition of the 32 kDa boar sperminogen showed significant differences from that of either boar proacrosin or ${\beta}-acrosin$, which signifies that 32 kDa sperminogen might not be a breakdown product of proacrosin-acrosin system and that the 32 kDa sperminogen is a different protein from proacrosin-acrosin system.

• Parasites, Hosts and Diseases
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• 제27권2호
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• pp.131-140
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• 1989

사람의 포충증과 유구낭미충증을 혈청학적으로 진단하는 데에는 교차반응이 상호 빈번히 일어나 임상상이나 기타 병력 등을 기초로 하여야 감별진단이 가능하다. 이 연구는 혈청학적 진단상 두가지 질환에서 교차반응을 일으키는 항원분획을 관찰하고 또 포충증과 유구낭미충증에 특이하게 반응하는 분획이 각 낭액중에 있는지를 관찰하기 위하여 실시하였다. 사우디·아라비아와 리비아에의 근무하고 귀국한 다음 발병한 포충증 환자 5예의 혈청과 유구낭미충증 환자 67예, 기타 간흡충증, 폐흡충증, 스파르가눔증, 무구조충감염자 89예 혈청을 포충과 유구낭미충 낭액을 항원으로 각각 효소면역측정법(ELISA)으로 특이항체가(IgG)를 측정하였다. 그 결과 포충 낭액에 대하여 포충증 혈청은 전례에서, 유구낭미충증 혈청은 49.3%에서, 기타 기생충증 혈청은 5.6%에서 양성반응을 보였다. 유구낭미충 낭액에 대하여 포충증 혈청은 전례가, 유구낭미충증 혈청은 55.1%가, 기타 기생충증 혈청은 12.3%가 양성반응을 나타내었다. 포충 낭액과 유구낭미충 낭액을 10∼15% linear gradient gel에서 SDS-PAGE를 실시한 바 포충 낭액에는 모두 19개 분획이, 유구낭미충 낭액에는 23개 분획이 나타났고 그 중 64, 35, 22, 7 kDa 분획이 두가지 낭액에 모두 나타나고 있었다. SDS-PAGE로 분리한 각낭액 분획을 항원으로 포충증과 유구낭미충증 환자 혈청을 반응시키고 반응분획을 발색시킨 바(면역얼룩법, immunoblot), 포충증 환자혈청은 포충 낭액의 145, 140, 135, 125, 117, 110, 100, 86, 64, 52, 45, 39, 35, 29, 24, 22, 17, 12 및 7 kDa분획에 반응하였고 유구낭미충 낭액에 대해서는 135, 110, 100, 86, 64, 45, 39, 35 및 24 kDa분획에 반응하였다. 또 유구낭미충증 환자혈청은 유구낭미충 낭액의 135, 130, 110, 105, 86, 72, 64, 57, 52, 45, 39, 55, 24, 22, 15, 10 및 7 kDa분획에 반응하였고 포충 낭액에는 135, 100, 86, 64, 52, 39, 35, 29 및 24 kDa 분획에 반응하였다. 이상의 결과에서 포충 및 유구낭미충 낭액에는 SDS-PAGE로 분리되는 분획중 135, 100, 86, 64, 39, 35 및 24 kDa 분획이 교차반응에 관여하는 공통항원 분획으로 판단되며 포충 낭액의 7 kDa 및 유구낭미충 낭액의 낭액의 15,10 및 7 kDa 분획은 특이항원 분획으로 생각된다.