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Updated: Aug 7, 2026

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
Published on: February 16, 2014
Whole genome phage display selects for proline-rich Boi polypeptides against Bem1p
Kirsten Hertveldt1, Johan Robben, Guido Volckaert
1Division of Gene Technology, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, Leuven B-3001, Belgium. kirsten.hertveldt@biw.kuleuven.be
Abstract:
Interaction selection by biopanning from a fragmented yeast proteome displayed on filamentous phage particles was successful in identifying proline-rich fragments of Boi1p and Boi2p. These proteins bind to the second "src homology region 3'' (SH3) domain of Bem1p, a protein of Saccharomyces cerevisiae involved in bud formation. Target Bem1p was a doubly-tagged recombinant, Bem1([Asn142-Ile551]), which strongly interacts in ELISA with a C-terminal 75 amino acids polypeptide from Cdc24p exposed on phage. The whole yeast genomic display library contained approximately 7.7 x 10(7) independent clones of sheared S. cerevisiae genomic DNA fused to a truncated M13 gene III. This study corroborates the value of fragmented-proteome display to identify strong and direct interacting protein modules.

