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Receptor-mediated endocytosis as a selection force to enrich bacteria expressing rhodostomin on their surface

  • Original Paper
  • Published:
Journal of Biomedical Science

Abstract

Previously, we developed a TraT display system to express snake venom rhodostomin (RHO), a disintegrin, on the external surface ofEscherichia coli [J Biomed Sci 6:64–70;1999]. To show a new potential use of the TraT display system, we employed a biotin labeling technique coupled with SDS-PAGE and flow cytometry analyses to further demonstrate and confirm the expression of TraT-RHO on theE. coli surface. We also showed that the expression of TraT-RHO on the cell surface not only facilitated the bacteria adhesion to BHK-21 cells but also induced bacterial internalization into BHK-21 cells. This feature allowed us to enrich the TraT-RHO expression bacteria about 10,000-fold starting with a mixture of TraT-RHO bacteria with β-galactosidase-positive bacteria in a ratio of 102:107 through four cycles of BHK-21 cell endocytosis and replating of engulfed bacteria on agar plates. We therefore suggest that the TraT display system can be applied to select out bacteria expressing a specific peptide sequence from a large population of display library through the process of receptor-mediated endocytosis and reamplification cycles.

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Authors and Affiliations

  1. Institute of Microbiology and Immunology, School of Life Science, National Yang-Ming University, Taipei, Taiwan 11221 (ROC)

    Hsin-Hou Chang, Ko-Nien Shih & Szecheng J. Lo PhD

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  1. Hsin-Hou Chang
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  2. Ko-Nien Shih
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  3. Szecheng J. Lo PhD
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Chang, HH., Shih, KN. & Lo, S.J. Receptor-mediated endocytosis as a selection force to enrich bacteria expressing rhodostomin on their surface. J Biomed Sci 7, 42–50 (2000). https://doi.org/10.1007/BF02255917

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  • DOI: https://doi.org/10.1007/BF02255917

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