• Title/Summary/Keyword: Western Blot

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Analysis of Placental Proteins in Somatic Cell Clone Recipient Cows

  • Woo, Jei-Hyun;Chung, Hak-Jae;Kim, Bong-Ki;Ko, Yeoung-Gyu;Kim, Jeom-Soon;Jung, Jin-Kwan;Chang, Won-Kyong
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.197-197
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    • 2004
  • The purpose of this experiment was to investigate the protein profiles in the placenta of Korean native cows(KNC) transferred cloned embryos and KNC artificially inseminated placental tissues were collected from the cows after cesarean section around parturition, and placental proteins were analyzed. Using two dimensional polyacrylamide gel eletrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. (omitted)

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Production of Recombinant Rotavirus Capsid Protein VP7 from Stably Transformed Drosophila melanogaster S2 Cells

  • Park, Jong-Hwa;Chang, Kyung-Hwa;Lee, Youn-Hyung;Kim, Hae-Yeong;Yang, Jai-Myung;Chung, In-Sik
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.563-568
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    • 2002
  • Stably transformed Drosophila melanogaster 52 cells producing recombinant VP7 were obtained, and recombinant VP7 expression was confirmed by Western blot analysis. The molecular weight of recombinant VP7 expressed in 52 cells was approximately 35.5 kDa, and 75% of the total VP7 produced was present in the medium. Recombinant VP7 contained N-linked glycosylated oligosaccharides. Aprotinin, leupeptin, and polyvinylpyrrolidone did not have any noticeable effect on recombinant VP7 production; however, DMSO and sodium butyrate increased its production by 120% and 60%, respectively.

The effects of Hominis Placenta extract on Melanin synthesis of B16 melanoma cells (자하거(紫何車) 약침액(藥針液)이 B16 melanoma 세포주의 멜라닌 합성에 미치는 영향)

  • Seo, Hyung-Sik
    • Journal of Pharmacopuncture
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    • v.9 no.1
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    • pp.75-82
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    • 2006
  • Objective : This research was carried out for the development of medicine for vitiligo treatment and focused on the effect of Hominis Placenta extract on Melanin synthesis of B16 melanoma cells. Methods : Acitivity of tyrosinase playing a vital role in synthesis of Melanin and the quantity of Melanin, which is the final product in cultured B16 melanoma cells, effects of Hominis Placenta extract were measured. Results: The results indicated that Hominis Placenta extract increased both the amount of Melanin and the activity of tyrosinase according to the concentration, and they also supported by western blot analysis. Conclusion : The results suggests that Hominis Placenta extract has an advantageous effect on the promotion of Melanin synthesis and will contribute to the development of vitiligo treatment through further related studies.

Characterization of Soluble Type Human Folate Receptor on Folate Binding and Transport (Soluble type의 human folate receptor의 folate 결합과 수송에 관한 특성)

  • Kim, Won-Sin
    • Toxicological Research
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    • v.14 no.3
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    • pp.371-377
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    • 1998
  • We have cloned a soluble type human folate receptor(hFR type${\gamma}$) from human thymus cDNA library using the PCR amplification technique. To examine whether hFR type${\gamma}$ has a folate transport activity, CHO cells were transfected with the pcDNAhFR${\gamma}$ expression plasmid, and the stable cell line CHO/hFR${\gamma}$ expressing a high level of the hFR type${\gamma}$ was identified by northern and western blot analysis. The CHO/hFR${\gamma}$ cells produced a [$H^3$]folic acid binding protein in the culture medium. However, we couldn't detect any cell surface [$H^3$] folic acid binding and transport activities. The growth of the CHO/hFR${\gamma}$ cells was more rapidly inhibited than the wild type CHO cells in the low concentration folic acid media. These observations indicate that although soluble type human folate receptor can bind [$H^3$]folate, it does not involve in folate transport.

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Specific Targeting of Fluorescein Isothiocyanate with Ep-CAM Antibody(Specific targeting of FITC with Ep-CAM Antibody)

  • Lee, Young-Tae;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.10 no.3
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    • pp.237-240
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    • 2003
  • The tetradecameric peptide (K47-K60) near the NH$_2$-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.

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Production of the recombinant fibronectin-binding protein of Staphylococcus aureus (Staphylococcus aureus의 재조합 fibronectin-binding protein의 생산)

  • kim, Doo;Cheong, Cha-ryong;Park, Hee-myong;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.37 no.4
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    • pp.875-882
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    • 1997
  • To produce the recombinant fibronectin-binding protein(FnBP) for development of subunit vaccine against Staphylococcus aureus. The fnbp gene was amplified from the chromosomal DNA of S aureus KNU 196 strain using the polymerase chain reaction, and cloned into pGEX-4T-2. Then, the recombinant FnBP fused with glutathione-S-transferase was produced in E coli, purified by affinity chromatography, and identified its antigenicity and immunogenicity by Western blot. The recombinant FnBP produced in this study is considered to have the same property of native FnBP purified from S aureus, and is expected to be useful as a candidate for S aureus subunit vaccine.

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Enhanced Secretion of Cell Wall Bound Enolase into Culture Medium by the sool-l Mutation of Saccharomyces cerevisiae

  • Kim, Ki-Hyun;Park, Hee-Moon
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.248-252
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    • 2004
  • In order to identify the protein(s) secreted into culture medium by the sool-l/retl-l mutation of Saccharomyces cerevisiae, proteins from the culture medium of cells grown at permissive (28$^{\circ}C$) and non-permissive temperatures (37$^{\circ}C$), were analyzed. Comparison of protein bands separated by SDS-PAGE identified a prominent band of 47-kDa band from a mutant grown at 37$^{\circ}C$. N-terminal amino acid sequencing of this 47-kDa protein showed high identity with enolases 1 and 2. Western blot analysis revealed that most of the cell wall-bound enolase was released into the culture medium of the mutant grown at 37$^{\circ}C$, some of which were separated as those with lower molecular weights. Our results, presented here, indicate the impairment of cell wall enolase biogenesis and assembly by the sool-l/retl-l mutation of S. cerevisiae.

Condensation of DNA by a Histone-like Protein in Escherichia coli

  • Kim, So-Youn;Hwang, Deog-Su
    • BMB Reports
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    • v.28 no.2
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    • pp.143-148
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    • 1995
  • In E. coli, chromosomal DNA associated with proteins is condensed into an organized structure known as nucleoid. Using a nitrocellulose filter binding assay to identify proteins forming nucleoid, a 21 kDa protein was purified from E. coli. The molecular weight of the purified protein was 21 kDa on SDS-polyactylamide gel electrophoresis and 24 kDa on gel permeation chromatography. A molecular weight of 21 kDa on SDS-polyacrylamide gel electrophoresis is unique among known proteins which are believed to be involved in the formation of nucleoid in E. coli. The 21 kDa protein nonspecifically binds to both double-stranded and single-stranded DNA. Sedimentation in a sucrose gradient revealed that the protein induced significant condensation of both supercoiled plasmid DNA and linear bacteriophage $\lambda$ DNA On the basis of quantitative Western-blot analysis, approximately 40,000 molecules of the protein were estimated to exist in an E. coli. The biochemical properties and cellular abundance of the 21 kDa protein suggest that this protein participates in the formation of nucleoid in E. coli.

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Expression Patterns of P21-Activated Kinase 2 by Sterne Spores on Human Macrophages (사람 대식세포에 스턴아포에 처리에 의한 P21-activated kinase 2 단백질의 발현양상)

  • Seo Gwi-Moon;Jung Kwang-Hwa;Kwak Hyun Jung;Kim Seong-Joo;Kim Ji-Chon;Chai Young-Gyu
    • Journal of the Korea Institute of Military Science and Technology
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    • v.7 no.4 s.19
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    • pp.87-90
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    • 2004
  • In order to elucidate the mechanism of infection on human macrophages, we peformed the 2-dimensional electrophoresis and the western blot analysis using the infected human macrophages with the spores of live and inactivated Sterne. We confirmed P21-activated kinase 2 protein which related to cell death(apoptosis) human macrophages at the early stage events. The inhibition of the P21-activated protein kinase 2 protein will be reduced apoptosis on infected human macrophages with Sterne spores.

Expression of a Yeast Superkiller Gene(SK13) in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 효모 Superkiller 유전자(SK13)의 발현)

  • ;Wickner, Reed B.
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.114-119
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    • 1990
  • A yeast chromosomal superkiller gene (SK13) was cloned and expressed in $ski3^{-}$ Saccharomyces cerevisiae strains. The gene was fused to the structural region of E. coli lacZ gene at its C-terminus in a yeast-E. coli shuttle vector, pSR605. The fused gene complemented $ski3^{-}$ strains with SK13 activity and the quantitative level of expression was measured as determined by assaying $\beta$-galactosidase activity. The SDS-polyacrylamide gel electrophoresis and the Western blot analysis of this fused protein showed the immuno-reacted bands with a protein of the estimated molecular size (ca.250Kd).

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