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Biotechnol Lett ; 43(8): 1649-1657, 2021 Aug.
Article in English | MEDLINE | ID: mdl-33934257

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.


Subject(s)
Cell Surface Display Techniques/methods , Plasmids/genetics , Recombinant Fusion Proteins , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Animals , Mice , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Viral Envelope Proteins/genetics , Viral Envelope Proteins/metabolism
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