Insertion of Peptide Coding Sequencing Sequence in pBAD BirA-eCPX-AP
1:32
Generation of a BirA Library
3:16
Selection of Bacteria Expressing Biotinylated Peptide
7:18
Quantification of Enrichment
8:26
Results: Model Selection with pBAD-BirA-eCPX-AP and pBAD-BirA-eCPX-AP(K10A)
10:00
Conclusion
副本
This protocol provides a highly effective bacterial display system for isolation of novel variants of BirA that allows for specific biotinylation of native proteins. The main advantage of the technique is that it selects BirA variants using high-a
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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.
