Titrating Avidity of Yeast-Displayed Proteins Using a Transcriptional Regulator.

ABSTRACT: Yeast surface display is a valuable tool for protein engineering and directed evolution; however, significant variability in the copy number (i.e., avidity) of displayed variants on the yeast cell wall complicates screening and selection campaigns. Here, we report an engineered titratable display platform that modulates the avidity of Aga2-fusion proteins on the yeast cell wall dependent on the concentration of the anhydrotetracycline (aTc) inducer. Our design is based on a genomic Aga1 gene copy and an episomal Aga2-fusion construct both under the control of an aTc-dependent transcriptional regulator that enables stoichiometric and titratable expression, secretion, and display of Aga2-fusion proteins. We demonstrate tunable display levels over 2-3 orders of magnitude for various model pro

SUBMITTER: Lopez-Morales J 

PROVIDER: S-EPMC9942200 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature