Reference: Piraine REA, et al. (2021) Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae. Biotechnol Lett 43(8):1649-1657

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Abstract


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.

Reference Type
Journal Article
Authors
Piraine REA, Gonçalves VS, Dos Santos Junior AG, Cunha RC, de Albuquerque PMM, Conrad NL, Leite FPL
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