| Autor: |
Pardavé-Alejandre, H. D., Alvarado-Yaah, J. E., Pompa-Mera, E. N., Muñoz-Medina, J. E., Sárquiz-Martínez, B., Santacruz-Tinoco, C. E., Manning-Cela, R. G., Ortíz-Navarrete, V., López-Macías, C., González-Bonilla, C. R. |
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| Zdroj: |
Biotechnology Letters; Mar2018, Vol. 40 Issue 3, p591-600, 10p |
| Abstrakt: |
Objectives: To display a recombinant avidin fused to the autotransporter ShdA to bind biotinylated molecules on the surface of Escherichia coli.Results: Two chimeric protein constructs containing avidin fused to the autotransporter ShdA were expressed on the surface of Escherichia coli DH5α. One fusion protein contained 476 amino acids of the ShdA α and β domains, whereas the second consisted of a 314 amino acid from α and truncated β domains. Protein production was verified by SDS-PAGE using an antibody to the molecular FLAG-tag. The surface display of the avidin-shdA fusion protein was confirmed by confocal microscopy and flow cytometry analysis, and the biotin-binding activity was evaluated by fluorescence microscopy and flow cytometry using biotin-4-fluorescein and biotinylated-ovalbumin (OVA).Conclusions: Expression of a recombinant avidin with biotin-binding activity on the surface of E. coli was achieved using the autotransporter ShdA. This system is an alternative to bind biotinylated molecules to E. coli. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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