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.
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Acknowledgements
The authors would like to thank CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) for promoting the scholarships offered to students involved in this study.
Funding
This study was funded by CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) and CNPQ (Conselho Nacional de Desenvolvimento Científico e Tecnológico).
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Piraine, R.E.A., Gonçalves, V.S., dos Santos Junior, A.G. et al. Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae. Biotechnol Lett 43, 1649–1657 (2021). https://doi.org/10.1007/s10529-021-03142-w
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DOI: https://doi.org/10.1007/s10529-021-03142-w