• 제목/요약/키워드: native- PAGE

검색결과 179건 처리시간 0.024초

Gel filtration에 의한 한방사선 인삼단백 분획의 정제 (Further Purification of Radioprotective Ginseng Protein Fraction by Gel Filtration)

  • 김춘미;박경애
    • 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.

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누에알의 배자발육에 따른 가용성 및 불용성 단백질의 변화 (Change in Soluble and Insoluble Proteins during Embryonic Development of the Silkworm, Bombyx mori)

  • 성주일
    • 한국잠사곤충학회지
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    • 제33권1호
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    • pp.1-5
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    • 1991
  • 누에알의 배자발율 단계별 가용성 단백질과 불용성 단백질에 대한 native-PAGE와 SDS-PAGE 분석을 하였다. 가용성 단백질의 대부분은 ESP, vitellin, 30K 단백질로 구성되어 있었고 배자발육 시기별 난황단백질의 변화는 native-PAGE와 SDS-PAGE에서 일치하였다. 배자발생의 후기에는 난황단백질의 분해산물로 보이는 수개의 peptides가 검출되었고 침산처리 후 1일된 알에서 1개의 특이한 단백질이 출현하였다. 누에알 마쇄물의 원심침전물에 대한 SDS-FAGE 분석결과 수종의 peptides가 배자발육의 진전과 함께 새로이 검출되었다.

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Properties of the Endonuclease Secreted by Human B Lymphoblastic IM9 Cells

  • Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • 제31권1호
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    • pp.106-110
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    • 1998
  • We have employed a DNA-native-polyacrylamide gel electrophoresis (DNA-native-PAGE) assay system to characterize the enzyme activity of the endonuclease secreted by human B lymphoblastic IM9 cells. Experimental results clearly demonstrated that the endonuclease activity of IM9 cell culture medium is distinct from that of DNase I in the DNA-native-PAGE assay system. Immunoprecipitation analysis using anti-DNase I antibody showed that the secreted endonuclease is not recognized by the antibody. The secreted endonuclease was estimated using supercoiled plasmid DNA as a substrate. The pH optimum required for the catalytic activity was determined to be in the range of pH 6.6-7.4. No significant difference in the endonuclease secretion was observed by stimulation of the IM9 cells with interferon-${\gamma}$ or interleukin-$1{\beta}$.

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Elctrophoretic Properties of Heat-Induced Bovine α-Lactalbumin

  • Lee, You-Ra;Hong, Youn-Ho
    • Preventive Nutrition and Food Science
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    • 제7권1호
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    • pp.48-51
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    • 2002
  • In order to study the reaction behavior of bovine holo- and apo-$\alpha$-lactalbumin ($\alpha$-La) during heat treatment at 65~10$0^{\circ}C$, the samples were analysed by first (ID)-and second-dimensional (2D) native-polyacrylamide gel electrophoresis (Native-PAGE) and sodium dodecylsulfate (SDS)-PAGE. When bolo-$\alpha$-La or apo- $\alpha$ -La were heated, they formed non-native, monomers, dimers and trimers. The apo-$\alpha$-La was more heat-sensitive than holo-$\alpha$-La. The monomers seemed to have the same composition as the native $\alpha$-La, but many of the disulfide bonds could be non-native.

Electrophoretic Behaviors of α-Lactalbumin and β-Lactoglobulin Mixtures Caused by Heat Treatment

  • Lee, You-Ra;Hong, Youn-Ho
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권7호
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    • pp.1041-1045
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    • 2003
  • In order to study the reaction behaviors of bovine $\alpha$-lactalbumin ($\alpha$-La), $\beta$-lactoglobulin ($\beta$-Lg), and their mixtures during heat treatment, samples were analyzed using native-polyacrylamide gel electrophoresis (Native-PAGE), sodium dodecylsulfate (SDS)-PAGE, and two-dimensional (2-D)-PAGE. The electrophoresis demonstrated that the loss of native-$\alpha$-La increased as temperature increased, and that the loss of apo-$\alpha$-La was slightly higher than that of holo-$\alpha$-La. The tests also showed that during heat treatment, a mixture of $\alpha$-La and $\beta$-Lg was less stable than $\alpha$-La alone. As such, it was assumed that $\beta$-Lg induced holo-$\alpha$-La to be less stable than apo-$\alpha$-La during heat treatment. The reaction behavior of $\alpha$-La (holo-, apo-form) during heat treatment showed similar patterns in the 2-D-PAGE electropherogram, but the mixture of $\alpha$-La and $\beta$-Lg created new bands. In particular, the results showed a greater loss of native $\alpha$-La in the holo-$\alpha$-La and $\beta$-Lg mixture than in the apo-$\alpha$-La and $\beta$-Lg mixture. Thus, it can be concluded that the holo-$\alpha$-La and $\beta$-Lg mixture was more intensively affected by heat treatment than other samples, and that free sulphydryl groups took part in the heat-induced denaturation.

한우(韓牛)에 있어서 Theileria sergenti의 항원성(抗原性)에 관(關)한 연구(硏究) (Study on the antigenicity of Theileria sergenti merozoite in Korean native cattle)

  • 백병걸;김병수;이재구
    • 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.

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PLGA 미립구로부터 PLGylated G-CSF의 서방성 방출 (Sustained Release of PLGylated G-CSF from PLGA Microsphere)

  • 정경환;임형권;이시욱;강관엽;박태관
    • KSBB Journal
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    • 제17권1호
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    • pp.33-37
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    • 2002
  • PLGA 미립구로부터 G-CSF의 방출 거동 양상을 향상 시키기 위하여 G-CSF를 분자량 5000인 methoxy polyethylene glycol-aldehyde로 PEGylation 시켰다. 대부분의 G-CSF는 mono-PEGylation 되었으며, 이를 SDS-PAGE, HPLC, 및 펩타이드 지도 분석을 통해 확인하였다. W/O/W 방법을 사용하여 G-CSF 및 PEGylated G-CSF의 PLGA 미립구를 제조하였으며, 이때 봉입율은 높은 상태였다. 미립구내로 더 많은 G-CSF를 봉입하기 위하여 액상의 G-CSF 및 PEGylated G-CSF을 농축하였고 native gel과 gel filtration 크로마토 그래피를 통하여 단백질이 안정함을 확인하였다. 이렇게 제조한 PLGA 봉입체의 in vitro 방출 거동을 조사한 결과 PEGylated G-CSF는 native G-CSF에 비하여 더 오랜 시간 동안 방출이 지속되었고 최대 방출량도 증가하였다.

한우 Lactoferrin 가수분해물의 항균 활성 (Antibacterial Activity of Enzymatic Hydrolyzates of Lactoferrin derived from Korean Native Cattle)

  • 이수원;양희진
    • 한국축산식품학회지
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    • 제18권4호
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    • pp.316-321
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    • 1998
  • In this study, we carried out to isolate lactoferrin from Korean native cattle and Holstein cow by batch extraction, ion exchange chromatography, gel filtration, and affinity chromatography. The purity of the isolated lactoferrin was higher than that of lactoferrin purchased from Sigma, when determined by SDS-PAGE and HPLC analysis. Antibacterial activity of E. coli O111 by Korean native cattle lactoferrin was lower than that of Holstein lactoferrin. A minimal inhibitory concentration(MIC) of Korean native cattle lactoferrin and Holstein lactoferrin was 2.75 mg/ ml and 1.5 mg/ml respectively. The lactoferrin hydrolyzate of Korean native cattle exhibited antimicrobial activity at 0.25 mg/ml, whereas that of Holstein cow exhibited antimicrobial activity at 0.12 mg/ml. The antibacterial potency of the hydrolyzate was at least tenfold greater than that of undigeated lactoferrin with strains tested. The effect of hydrolyzate was bactericidal as indicated by rapid loss of viability of E. coli O111.

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Bacillus subtilis A405 균주가 생성하는 내열성 항균 peptide의 특성 검정 (Characterization of a heat-resistant antimicrobial peptide secreted by Bacillus subtilis A405)

  • 구본성;이승범;윤상홍;송재경;정대성;변명옥;류진창
    • 농약과학회지
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    • 제2권3호
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    • pp.28-35
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    • 1998
  • 전국에서 분리한 세균으로부터 항균력이 있는 길항세균들을 먼저 분리한 다음 그중 배양배지 속으로 강력한 항균력을 나타내는 peptide를 분비하는 A405균주를 선발, Biolog System 및 rDNA 염기서열을 이용하여 Bacillus subtilis로 동정하고 이 균주가 생성하는 항균 peptide의 항균력을 조사한 바 Botrytis cinerea, Cercospora sp., Fusarium oxysporum, Penicillium digitatum, Celletotrichum gloeosporioides, Rhizoctonia solani, Pythium ultimum, Pyricularia oryzae등의 식물병원균과 Escherichia coli, Pseudomonas spp. 등의 세균 및 효모인 Candida albicans의 성장도 억제하는 강력한 항균력을 보였다. 또한 이 균주가 생산하는 항균 peptide는 알콜, 아세톤과 같은 유기용매에 내성을 가지고 있었으며 proteinase K와 phenol 처리시에는 항균력이 감소되거나 소실되는 특성을 가지고 있었다. 이 항균 peptide를 SDS-PAGE, Native-PAGE 및 Tris-Tricine 전기영동으로 분자량이 3kDa인 작은 peptide임을 확인하였고 aspartic acid, glycine, serine, glutamine, valine, leucine, isoleucine, proline, tyrocine 등의 9가지 아미노산으로 구성되어 있음을 Amino Acid Analyzer로 확인하였으며 냉동건조한 항균 peptide 50 ug/ml을 체리토마토에 처리하여 과일의 선도유지를 조사한 결과 대조구에 비하여 월등한 선도유지 효과가 있음을 입증하였다.

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Comparison of Postmortem Proteolysis between Breast and Leg Muscles in Chiayi Native Chickens

  • Cha, Shih-Ting;Tseng, Tsai-Fuh;Ho, Sy-Shyan;Chou, Rong-Ghi R.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권5호
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    • pp.721-724
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    • 2002
  • Postmortem Proteolysis of breast (BM) and leg (LM) muscles in Chiayi native chickens at $5^{\circ}C$ were compared. Myofibrils were purified from BM and LM samples that were randomly taken from carcasses after 0, 1, 3, 7 and 14 days of storage at $5^{\circ}C$. Fragmentation of myofibrils were determined, and degradation of myofibrillar proteins were analyzed by the SDS-PAGE and western blots. The results showed that myofibril fragmentation index (MFI) was significantly (p<0.05) higher in BM than in LM samples. Disappearance of titin and nebulin and appearance of the 30 kDa component were more rapidly as seen on SDS-PAGE in BM than in LM samples. Western blots labeled with a monoclonal antibody to desmin also demonstrated that desmin degraded more quickly in BM samples. Our data suggested that postmortem proteolysis occurred more rapidly in breast muscles in Chiayi native chickens.