A Protein-Engineered, Enhanced Yeast Display Platform for Rapid Evolution of Challenging Targets.

ABSTRACT: Here, we enhanced the popular yeast display method by multiple rounds of DNA and protein engineering. We introduced surface exposure-tailored reporters, eUnaG2 and DnbALFA, creating a new platform of C and N terminal fusion vectors. The optimization of eUnaG2 resulted in five times brighter fluorescence and 10 °C increased thermostability than UnaG. The optimized DnbALFA has 10-fold the level of expression of the starting protein. Following this, different plasmids were developed to create a complex platform allowing a broad range of protein expression organizations and labeling strategies. Our platform showed up to five times better separation between nonexpressing and expressing cells compared with traditional pCTcon2 and c-myc labeling, allowing for fewer rounds of selection and achievi

SUBMITTER: Zahradnik J 

PROVIDER: S-EPMC8689690 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature