• 대한수의학회지
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• 제38권2호
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• pp.328-335
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• 1998

Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8.

• Journal of Plant Biology
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• 제17권4호
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• pp.143-156
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• 1974

The purpose of this experiment was to study one side of germination physiology based on that protein profiles and protease relating to protein metabolism, that peroxidase, catalase, $\alpha$-amylase, $\beta$-amylase, and malate dehydrogenase involved in the carbohydrate metabolism of seed germination. All these experiments were divided into the two groups with and without acetone treatment, and were carried out. The protein bands of each germinating stage between the groups treated with and without acetone showed certain basic pattern in polyacrylamide gel disc electrophoresis. However, there was a little difference in the number of protein band, optical density, and migration velocity between two groups. The isozyme bands of peroxidase, and catalase between two groups in polyacrylamide gel disc electrophoresis did not show the numeral difference, but the optical density of certain germinating stage treated with acetone was higher than the group untreated with it and it showed their enzyme activity. The $\alpha$-amylase and $\beta$-amylase activities which involved in starch metabolism of seed germination were higher in the treated group than the other. On one hand, the protease activity of hydrolase occurred in the seeds for germination was also higher, more or less in the treated group than in the other. The isozyme band pattern of malate dehydrogenase in TCA cycle of energy metabolism pathway was very different between two groups growing for 72 hours with and without acetone treatment in cellulose acetate electrophoresis. It indicated that two isozyme bands of malate dehydrogenase was high. Consequently these experimental results mentioned above indicated that acetone treatment before sowing had an effect on dissolving certain complexed lipid substance involved in the seed coats, the activity of carbohydrate hydrolase increased with water absorption which was most comfortable in its germination, dissolved glycerin and fatty acid became certain energy source, and they stimulated the acceleration of respiration metabolism.

• 한국해양바이오학회지
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• 제2권1호
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• pp.68-72
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• 2007

넙치의 혈청 단백질체를 연구하기 위해 본 연구에서는 이차원전기영동 분석 시의 기본조건을 확립하였다. 넙치의 혈청단백질을 이차원전기영동 법으로 분석하기 위해서는 TCA 침전을 이용하여 혈청 내에 고농도로 존재하는 이온물질을 제거하는 전처리과 정이 반드시 요구되었다. 또한 분석 단백질의 농도는 $30{\mu}g$이 적당하였다. 넙치 10개체의 혈청단백질을 이용하여 총 51회의 이차원전기영동을 수행한 결과 1,8207개의 단백질로 이루어진 넙치 혈청단백질 표준 지도를 작성할 수 있었다.

• 농업과학연구
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• 제45권4호
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• pp.635-643
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• 2018

Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

• 한국산림과학회지
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• 제50권1호
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• pp.5-9
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• 1980

Polyacrylamide gel 전기영동법(電気泳動法)을 이용(利用)하여, 슬래쉬 소나무 (접목묘(接木苗), 20년생(年生))의 화아원기형성기간(花芽原基形成期間) 동안에 자화다량(雌花多量) 개화개체(開花個体)와 소량(少量) 개화개체간(開花個体間)에 정아(頂芽)의 peroxidase 동위효소(同位酵素)의 변화(変化)를 비교(比較)하였다. 정아(頂芽)에서 모두 9개의 band를 관측(観察)할 수 있었으며, 두 개체간(個体間)에 동위효소(同位酵素)의 양적(量的)인 차이(差異)는 미세(溦細)하였으나, 제오(第五) band는 다량(多量) 개화개체(開花個体)에서 대부분(大部分) 발견(発見)되었다. 제오(第五) band의 동위효소(同位酵素)는 자화원기형성(雌花原基形成)에 직접(直接) 관병(関聠)되는 것 같이 생각된다.

• Journal of Plant Biotechnology
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• 제31권4호
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• pp.319-323
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• 2004

Euphorbia lathyris의 유관세포로부터 분비되는 수용성 latex단백질을 10% SDS-polyacrylamide 전기영동으로 분리하여 특징적으로 잘 나타나고 있는 ELp65, ELp55, ELp43, ELp32 그리고 ELp23 등의 주요단백질을 확인하였다. 이들 latex 주요단백질들 중에서 ELp65는 ammonium sulfate 침전, gel permeation chromatography 그리고 ion exchange chromatography 등의 방법으로 순수 분리하였는데 ELp65의 N-terminal amino acid 서열분석에 의하면 이는 토마토의 p69a subtilisin-like pretense의 mature peptide의 앞부분과 강한 유사성을 지니고 있었으며 식물방어와 관련된 기능이 제시되었다. PCR증폭에 의하여 클로닝된 토마토의 p69a DNA를 probe로 이용하여 Southern blot hybridization을 수행한 결과 E. lathyris genome은 토마토의 subtilisin-like proteases와 유사한 정보를 지닐 수도 있는 3-5 유전자들로 구성된 gene family가 분석되었다.

• 한국동물학회지
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• 제24권1호
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• pp.27-37
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• 1981

노랑초파리 (Drosophila melanogaster) 난자형성단계에 따른 난소와, 수정 후 발생에 EK른 발생배, 유충 그리고 번데기의 단백질양상을 polyacrylamide gel 전기영동법으로 조사하였다. 초파리 난자형성단계에 따른 난소의 단백질양상과, 수정 후 발생단계에 따른 단백질양상은 각 단계가 진행됨에 따라 전기영동상에서 band의 차이로 나타나는 질적인 면과 상대적인 양에 있어서 다양한 변화를 보여 주었다. 특히 난자형성 초기단계에서는 3개로 나타났던 난황단백질이 후기에는 한 개의 단백질로 전환해가는 현상을 나타내었다. 그리고 번데기시기까지의 단백질양상은 알의 시기에서 유충시기로의 전환점과 2령기에서 3령기로 전환되는 점에서 큰 변화를 보여 주었다.

• 한국식품영양과학회지
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• 제18권3호
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• pp.279-286
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• 1989

Alkaline protease 생성능이 강한 Aspergillus fumigatus을 토양에서 분리동정하고, 효소생산조건을 구명한 결과 $30^{\circ}C$에서 3일간 배양하였을 때 최고활성을 나타냈으며 생산된 조효소를 황산암모늄염석, Sephadex G-25, G-150 gel filteration과 DEAE-Cellulose컬럼 chromatography로 정제하여 수율 6.4%, 정제정도 86.13배의 효소를 얻었고 polyacrylamide gel 전기영동에 의해 단일밴드인 것을 확인하였다. 정제효소의 분자량은 63000정도 였으며, 17종의 아미노산으로 구성되어 있고, 그 중 glycine과 glutamic acid가 가장 많고 methionine과 cystine이 가장 적었다.

• 한국미생물·생명공학회지
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• 제9권4호
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• pp.213-218
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• 1981

순수분리정제된 $\beta$-galactosidase의 분자량은 Sephadex G-200 gel filtration법에서 130,000이며 SDS-polyacrylamide gel electrophoresis에서 130,000외에 70,000이 확인되어 이 효소는 70,000인 2개의 subunit로 구성되어 있다고 판단되었다. 효소의 안정pH는 4.5~7.0이며 효소활성의 최적 pH는 4.7 최적온도는 5$0^{\circ}C$였다. 효소활성 및 안정제로 1가, 2가 금속ion을 필요로 하지않았으며 C $u^{++}$ 1mM에서 59%, galactose 100mM에서 48% 저해되었다. 5% lactose용액, 시유 및 10% skim milk 용액에 이 정제된 $\beta$-galactosidase 10units/$m\ell$를 사용하여 5$0^{\circ}C$에서 4시간 동안 반응시켰을 때 lactose가 각각 69.5%. 88.7% 및 72.6%가 glucose와galactose로 전환된 결과를 얻었다.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.39-45
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• 2002

Activity staining on the native polyacrylamide gel electrophoresis (PAGE) of a cell-free extract of Rhodococcus sp. TK6, grown in media containing alcohols as the carbon source, revealed at least seven isozyme bands, which were identified as alcohol dehydrogenases that oxidize cyclohexanol to cyclohexanone. Among the alcohol dehydrogenases, cyclohexanol dehydrogenase II (CDH II), which is the major enzyme involved in the oxidation of cyclohexanol, was purified to homogeneity. The molecular mass of the CDH II was determined to be 60 kDa by gel filtration, while the molecular mass of each subunit was estimated to be 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The CDH II was unstable in acidic and basic pHs, and rapidly inactivated at temperatures above $40^{\circ}C$ . The CDH II activity was enhanced by the addition of divalent metal ions, like $Ba^2+\;and\;Mg^{2+}$. The purified enzyme catalyzed the oxidation of a broad range of alcohols, including cyclohexanol, trans-cyclohexane-1,2-diol, trans-cyclopentane-l,2-diol, cyclopentanol, and hexane-1,2-diol. The $K_m$ values of the CDH II for cyclohexanol, trans-cyclohexane-l,2-diol, cyclopentanol, trans-cyclopentane-l,2-diol, and hexane-l,2-diol were 1.7, 2.8, 14.2, 13.7, and 13.5 mM, respectively. The CDH II would appear to be a major alcohol dehydrogenase for the oxidation of cyclohexanol. The N-terminal sequence of the CDH II was determined to be TVAHVTGAARGIGRA. Furthermore, based on a comparison of the determined sequence with other short chain alcohol dehydrogenases, the purified CDH II was suggested to be a new enzyme.