Related Experiment Video
Updated: Nov 7, 2025
Protein Engineering by Yeast Surface Display
Published on: November 29, 2024
Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae
Renan Eugênio Araujo Piraine1, Vitória Sequeira Gonçalves1, Alceu Gonçalves Dos Santos Junior1
1Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Objectives:
Develop a Cell Surface Display system in Saccharomyces cerevisiae, based on the construction of an expression cassette for pYES2 plasmid.
Results:
The construction of an expression cassette containing the α-factor signal peptide and the C-terminal portion of the α-agglutinin protein was made and its sequence inserted into a plasmid named pYES2/gDαAgglutinin. The construction allows surface display of bovine herpesvirus type 5 (BoHV-5) glycoprotein D (gD) on S. cerevisiae BY4741 strain. Recombinant protein expression was confirmed by dot blot, and indirect immunofluorescence using monoclonal anti-histidine antibodies and polyclonal antibodies from mice experimentally vaccinated with a recombinant gD.
Conclusions:
These results demonstrate that the approach and plasmid used represent not only an effective system for immobilizing proteins on the yeast cell surface, as well as a platform for immunobiologicals development.
