• Biomolecules & Therapeutics
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• 제9권1호
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• pp.7-14
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• 2001

The chicken meat has been reported as one of the food causing allergic reactions predominantly to Korean. At present, several in vitro tests for immunoglobulinG (IgG)-mediated as well as IgE-mediated food allergy are available. 13 clinically chicken meat-allergic patients were investigated together with 4control subjects for identification of chicken meat-specific reactivity by ELISA. Also, protein profile and IgE, IgGtotal and IgG4-reacting allergens were detected by means of sodium dodecyl sulfate-polyacrylamide gel electro-phoresis (SDS-PAGE)and immunoblotting. Chicken meat extracts were prepared as raw, heated, heat and simulated gastric fluid (SGF) treated samples to characterize the stability of allergen to physicochemical treatment. SDS-PAGE revealed 9~200 kDa bands. And in immunoblotting 7 sera were identified most major bands between 10 and 78 kDa. In case of IgE, six proteins (17, 26, 35, 40, 78 kDa) were predominant in heat-treated extract, and the one (35 kDa) was present in SGF-treated preparations. In case of IgG$_{total}$ and IgG4, most of them showed a patters simmilar to IgE. There were significant differences (P<0.05) in IgE, IgG$_{total}$ , IgG4 Abs to chicken meat between the allergic and control subjects in ELISA. In addition, the concentration of IgG4Abs in the challenge-positive subjects was significantly higher than that of control subjects. It is considered that the specific IgE response to chicken meat was rarely prevalent to Koreans. However, the specific IgG4 response play an important role in the development of allergic symptoms.

• 한국작물학회지
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• 제43권4호
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• pp.228-233
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• 1998

Seed storage proteins have been used for studying biochemical genetics and end-use quality aspects. We conducted enzyme-linked immunosorbent assay (ELISA) and one-dimensional SDS-PAGE (1D SDS-PAGE) to evaluate different cereal crop species and Korean wheat lines for rye secalin proteins. The antisecalin antibody showed consistent specificity for rye secalin with little cross-reactivity to gliadins. Immunological cross-reactivities measured by the ELISA technique using competition assay showed significant differences of absorbance among rye, triticale, wheat-rye translocated wheat and non-translocated wheat. The absorbance values were lowest in rye followed by triticale, translocated wheat and non-translocated wheat. The ELISA for discrimination of wheat-rye translocation on the basis of antigen-antibody reactivity showed that none of the Korean wheat lines possessed 1RS and secalin proteins. The competitive ELISA experiment demonstrated specific determination for secalin that was originated from rye chromosomal parts. The result of 1D SDS-PAGE for identifying rye secalin subunits showed all three rye specific secalin protein subunits (75 KDa, 45 KDa, and 40 KDa) for rye and triticale, and 1RS specific secalins (45 KDa and 40 KDa) for 1AL/1RS and 1BL/1RS translocated wheats. All Korean wheats were lacking 1RS of rye chromosome and secalin.

• 한국어병학회지
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• 제5권2호
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• pp.87-92
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• 1992

넙치의 혈청, 체표점액 및 각 조직으로부터 lysozyme의 분리와 정제를 Chitin-coated Cellulose column 크로마토그라피를 사용하여 분리하였다. 15% SDS-PAGE의 전기 영동상을 조사한 결과 넙치의 신장, 유문수, 아가미 점액 및 혈청으로부터 분리된 lysozyme은 동일한 분자량(약 14000)을 나타내었다. 넙치의 방어기작으로서 lysozyme의 역할을 밝히기 위해 그람양성 세균 2주, 그람음성 세균 5주에 대하여 넙치 신장 유래 lysozyme의 정균작용을 조사한 결과 그람 양성 2주 그람 음성 세균 2주에 대하여 정균작용을 나타내었다. 이것은 넙치 유래의 lysozyme은 그람 양성 세균뿐만 아니라 그람 음성 세균에 대해서는 생체 방어 작용을 수행하는 것으로 고려된다.

• 미생물학회지
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• 제36권1호
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• pp.14-19
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• 2000

Penicillium oxalicum(HCLF-34)의 세포외 분비효소로부터 ultrafilration, gel filtration chromatograph와 anion exchange chromatography법을 이용하여 남조세균(Anabaena cylindrica) 분해효소를 분리하였다. 이 효소의 분자량은 약 22 kDa이며, renaturation SDS-PAGE에서 monomer로서 남조세균 분해 활성을 갖는다. 아미노산 서열은 N-말단부터$NH_(2)$-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, 13개의 아미노산을 분석하였으며, 분석된 아미노산의 homology를 조사한 결과 aspergillopepsin II precursor(acid protease A)와 13개의 아미노산 중 11개(84%)의 유사도를 나타내었고, acid proteinase EapC precursor과 13개 중 10개(81%)의 유사도를 나타내었다.

• 대한수의학회지
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• 제30권2호
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• pp.223-229
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• 1990

A splenectomized 5-month-old calf was inoculated with cryopreserved Theileria sergenti infected blood originated from naturally infected Korean native cattle in Chonbuk district. At peak parasitemia (40.1%), blood was collected, washed, lysed and then the T sergenti merozoite was isolated by differential centrifugation. Antigenic profile of isolated T sergenti organism was analized by SDS-PAGE and western blotting techniques. Coomassie blue stained SDS-PAGE gel revealed at least twelve protein bands of approximately 14Kd, 28Kd, 30Kd, 34Kd, 36Kd, 38Kd, 41Kd, 56Kd, 66Kd, 72Kd, 97Kd and 116Kd in the merozoite homogenate. In western blot, although T sergenti antigen recognized by specific anti-T sergenti antibodies demonstrated 28Kd, 30Kd, 38Kd, 56Kd, 58Kd, 66Kd, 97Kd and 116Kd proteins. False positive reactions were also observed in normal bovine serum with T sergenti and normal erythrocytic antigens. Therefore, predominant proteins of T sergenti merozoite antigen were found to be 28Kd, 30Kd, and 41Kd proteins of molecular weights. On going studies we will analyze the relative importance of those antigens for immunity of T sergenti in Korean native cattle.

• 한국미생물·생명공학회지
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• 제19권3호
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• pp.303-307
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• 1991

B.thuringiensis subsp. darmstadiensis 73E10-2의 내독소에 존재하는 hemolysin이 Sephadex G-100 gel filtration과 DEAE-cellulose ion exchange column chromatography에 의해 정제되었으며, 그 순도는 SDS-PAGE와 Ouchterlony test로 확인하였다. 그 결과 정제된 hemolysin의 분자량은 64KDa 의 단백질 이었으며, in vivo 상태에서는 전혀 모기유충에 독작용을 나타내지 않았다는 점이 28KDa 단백질의 차이가 있었다. 또한 정제된 hemolysin과 B.thuringiensis subsp. israelensis의 내독소를 효소항체법으로 검토해 본 결과 양단백질 사이에는 면역학적으로 전혀 상관이 없었다.

• 농업과학연구
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• 제24권2호
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• pp.121-125
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• 1997

베트남으로부터 수집된 총 437 벼 지역종을 사용하여 총 종자 단백질을 분석하기 위하여 SDS-PAGE와 phenol 반응을 수행하였다. 세 가지 다른 형태의 glutelin a subunit이 검출되었다. 60kDa의 분자량인 wx 단백질 수준은 세 그룹으로 분리되었는데, 이는 non-glutelin, intermediate, 그리고 glutelin 전분 유형이다 종자 저장 단백질과 wx 단백질의 변이에 의한 실험결과 지역종은 7그룹으로 구분된다. Glutelin a subunit의 A와 B형의 빈도분포는 벼 지역종이 수집된 위도에 따라서 변하였다. 페놀 반응에 대한 지리학적인 구배가 검출되었다.

• 한국식품영양과학회지
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• 제26권6호
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• pp.1116-1121
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• 1997

Effects of gamma irradiation onf the activity and the properties(amino acid compositions, in vitro digestibility and SDS-PAGE pattern) of proteolytic enzymes were investigated. The proteolytic activity of soluble human serine protease, enzyme in kiwi and pineapple decreased 10% and 30~65% by 5 kGy and 30 kGy, respectively. In dried pancreatin and lysozyme, the proteolytic and antimicrobial activities decreased 6~14% and 10~20% by 5kGy and 40kGy, respectively. The analysis of above 10kGy-irradiated soluble human serine protease by SDS-PAGE revealed radiolysis of the enzyme into protein or peptides of lower molecular weights. The irradiation of skim milk, hammastein casein, and lysozyme up to 40kGy had no deleterious effect on either the in vitro digestibility or amino acid compositions.

• Restorative Dentistry and Endodontics
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• 제22권2호
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• pp.693-701
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• 1997

In 1992, Prevotella intermedia was shown to be comprised of another spoecies now known as Prevotella nigrescens. Strain ATCC 33563 is now designated the type strain of P. nigrescens while strain ATCC 25611 is remains the type strain of P. intermedia. The purpose of this study was to find the differences in protein profiles of P. intermedia and P. nigrescens, using sodium dodecyl sulfate polyacrylamide gel electrophoresis, which can be used for differentiation of those two species. A partial amino acid sequence of the 18.6 kDa protein band, which was specific in P. nigrescens, was also determined. The cellular proteins were extracted from the cell pellets of pure cultures of P. intermedia. and P. nigrescens by either sonication or being shaken continuously for 20 min at $21^{\circ}C$ with 1 % SDS or being boiled for 3 min with 1 % SDS. SDS-PAGE was performed according to the method of laemmli using either 12% (w/v) gels or 18% (w/v) gels. Results were as follows ; 1. The similar electrophoretic protein profiles were shown by 3 cellular protein extraction methods for each strain. (Fig. 1 and 2) 2. the 18.6 kDa band which was specific only in P. nigrescens could be used for the differentiation of P. intermedia. and P. nigrescens. (Fig. 1 and 2, Table 1) 3. A total of 4 different tryptic fragments from the 18.6 kDa protein were sequenced. the resulting amino acid sequences were fragment 1.GNPVNIGGEW, 2.FNVVR, 3.NYLT-VAPY, and 4.GGDNVTTYQVLPEIGYN. By comparison to the sequences of known proteins in the Swiss-Prot database and PIR database. 90 % matching between fragment 1 and serine hydroxymethyl transferase(P24060) in the Swiss-Prot, and 90% matching between fragment 1 and glycine hydroxymethyl transferase(S15203) in the PIR were shown, but the identity and function of the 18.6 kDa protein remains unknown.

• 한국식물병리학회지
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• 제11권4호
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• pp.312-317
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• 1995

The polygalacturonase (PG) production in rotten apples by Botryosphaeria dothidea was purified by using gel filtration and ion exchange column chromatography, and the biochemical characters of PG were investigated. The purified PG appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with approximate molecular weight of 49 kilodalton (kDa). The molecular weight was equal to the native molecular weight estimated by gel filtration. The Km and Vmax values of PG were 0.51 mg/ml and 90.9 $\mu$M/min/ml, respectively. Optimum pH was 4.0~5.0, and the PG activity was stable from pH 5.0~10.0. Optimum temperature of the enzyme activity was 4$0^{\circ}C$. The PG activity was relatively stable at 2$0^{\circ}C$, but it was reduced 45% at 4$0^{\circ}C$ and completely inactivated at 8$0^{\circ}C$. The PG activity was considerably inhibited by Cu2+, Zn2+, SDS and EDTA, whereas it was not effected by Ca2+, K+, Mg2+ or Na+ ions.