Autodisplay of an avidin with biotin-binding activity on the surface of Escherichia coli.
Biotechnol Lett
; 40(3): 591-600, 2018 Mar.
Article
em En
| MEDLINE
| ID: mdl-29357100
ABSTRACT
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 ß domains, whereas the second consisted of a 314 amino acid from α and truncated ß 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.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas da Membrana Bacteriana Externa
/
Proteínas Recombinantes de Fusão
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Avidina
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Escherichia coli
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Proteínas de Membrana
Idioma:
En
Revista:
Biotechnol Lett
Ano de publicação:
2018
Tipo de documento:
Article