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- Title
Receptor-Mediated Endocytosis as a Selection Force to Enrich Bacteria Expressing Rhodostomin on Their Surface.
- Authors
Chang, Hsin-Hou; Shih, Ko-Nien; Lo, Szecheng J.
- Abstract
Previously, we developed a TraT display system to express snake venom rhodostomin (RHO), a disintegrin, on the external surface of Escherichia 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 the E. 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 10[sup 2] :10[sup 7] 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.Copyright © 2000 National Science Council, ROC and S. Karger AG, Basel
- Subjects
ENDOCYTOSIS; BACTERIA; ABSORPTION (Physiology); CELL physiology; PROKARYOTES
- Publication
Journal of Biomedical Science, 2000, Vol 7, Issue 1, p42
- ISSN
1021-7770
- Publication type
Article
- DOI
10.1007/BF02255917