• Title, Summary, Keyword: native-PAGE

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

  • 김춘미;박경애
    • Journal of Ginseng Research
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    • v.13 no.2
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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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Change in Soluble and Insoluble Proteins during Embryonic Development of the Silkworm, Bombyx mori (누에알의 배자발육에 따른 가용성 및 불용성 단백질의 변화)

  • 성주일
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.1-5
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    • 1991
  • PBS soluble- and insoluble-extracts form silkworm eggs were analysed by native-PAGE and SDS-PAGE. During the embroyonic development, soluble proteins which are mainly composed of ESP, vitellin and 30K proteins showed similar degradation pattern in both electrophoretic analyses. Several peptides which seemed to be intermediated forms of yolk proteins were detected in latter part of embryogenesis, while a protein band newly appeared in one day elapsed after acid treatment. When insoluble extracts from silkworm eggs were analysed with SDS-PAGE, several peptides were detected at the later stages of the embryonic development, and newly hatched larvae. These peptides are considered to be structure proteins for embryonic morphogenesis.

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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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    • v.31 no.1
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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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    • v.7 no.1
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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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    • v.16 no.7
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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.

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

  • Baek, Byeong-kirl;Kim, Byeong-su;Rhee, Jae-ku
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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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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Sustained Release of PLGylated G-CSF from PLGA Microsphere (PLGA 미립구로부터 PLGylated G-CSF의 서방성 방출)

  • 정경환;임형권;이시욱;강관엽;박태관
    • KSBB Journal
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    • v.17 no.1
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    • pp.33-37
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    • 2002
  • To improve in vitro release kinetic of G-CSF in PLGA microsphere, G-CSF was PEGylated with methoxy polyethylene glycol-aldehyde (mPEG-aldehyde, MW 5000). The majority of G-CSF was mono-PEGylated and it was characterized using SDS-PAGE, HPLC, and peptide mapping. The PLGA microencapsulation with the native, or PEGylated G-CSF was performed using W/O/w method, where the encapsulation efficiency was high. For the high loading of G-CSF to microsphere, G-CSF and PEGylated G-CSF were concentrated and then verified the protein stability using native gel and gel filtration chromatography. In comparison with native G-CSF, PEGylated G-CSF was released during the extended period and its maximum amount of released G-CSF was also increased.

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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    • v.15 no.5
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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.

Comparision of Mineral, Hydroxy Methyl Furfural Content and SDS-PAGE Pattern of Proteins in Different Honeys (다양한 꿀에 함유된 무기물 조성, Hydroxy Methyl Furfural 함량 및 꿀 단백질의 전기영동 패턴 비교)

  • Jung, Mi-Ea;Kim, Cheon-Jei;Paik, Hyun-Dong;Oh, Jae-Wook;Lee, Si-Kyung
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.241-249
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    • 2011
  • This study was conducted to analyze ash content, mineral composition, hydroxy methyl furfural (HMF) content, stable carbon isotope ratio, and SDS-polyacrylamide gel electrophoresis patterns to investigate the quality characteristics of various honeys harvested from different sources and to identify differences useful for distinguishing honey sources. Ash content was 0.046-0.012% in acacia honey, 0.565-1.318% in chestnut honey, 0.06-0.582% in polyfloral honey, and 0.237-0.893% in native bee honey. Potassium content was high in order of chestnut honey>native bee honey>polyfloral honey>acacia honey. The Na/K ratio was 0.92-1.97 in acacia honey, 0.02-1.59 in chestnut honey, 0.02-5.30 in polyfloral honey, and 0.22-0.51 in native bee honey. The HMF content was 9.60-12.85, 10.15-25.75, 9.7-33.5, and 6.25-21.5 mg/kg in acacia, chestnut, native bee, and polyfloral honeys, respectively. HMF content was the highest in native bee honey. A 59 kDa protein band was revealed in all samples by SDS-PAGE analysis. Protein bands of 32.1, 31.9, and 33.5 kDa were revealed in some chestnut honeys, and protein bands of 32.3 and 32.5 kDa were shown in native bee honeys. A protein band of 72 kDa was also confirmed in some chestnut honeys.

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

  • 이수원;양희진
    • Food Science of Animal Resources
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    • v.18 no.4
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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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