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Immunology and Infection

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Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Article DOI: 10.3791/60266-v • 10:43 min • October 3rd, 2019
October 3rd, 2019 •
1, 1,2, 1
1Department of Molecular Medicine, University of Southern Denmark, 2Department of Nephrology, Odense University Hospital

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Summary
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Here we present a method to select for novel variants of the E. coli biotin-protein ligase BirA that biotinylates a specific target peptide. The protocol describes the construction of a plasmid for the bacterial display of the target peptide, generation of a BirA library, selection and characterization of BirA variants.

Tags

Bacterial Peptide Display Selection Of Biotinylating Enzymes BirA Variants Streptavidin Selection Efficient Selection Process PCR Reaction Amplification Agarose Gel Mutant Megaprimers PBAD-BirA-eCPX Target Peptide Sequence
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