Abstract
Develop a Cell Surface Display system in Saccharomyces cerevisiae, based on the construction of an expression cassette for pYES2 plasmid. 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. 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.
Highlights
The construction of an expression cassette containing the α-factor signal peptide and the Cterminal portion of the α-agglutinin protein was made and its sequence inserted into a plasmid named pYES2/gDαAgglutinin, allowing cell surface display of bovine herpesvirus type 5 (BoHV-5) glycoprotein D on S. cerevisiae BY4741 strain
These results demonstrate that the approach and plasmid used represent an effective system for immobilizing proteins on the yeast cell surface, as well as a platform for immunobiologicals development
Cell Surface Display is a technique developed for recombinant protein expression in heterologous systems, such as bacterial, insect and yeast cells (Bertrand et al 2016)
Summary
Cell Surface Display is a technique developed for recombinant protein expression in heterologous systems, such as bacterial, insect and yeast cells (Bertrand et al 2016). The commercial availability of plasmid vectors for Yeast Surface Display (YSD) is generally limited to options that depend on the a-agglutinin anchoring system (Yang et al 2019), new plasmids able of heterologous proteins anchoring should be sought as possible alternatives to implement this method. The technique success depends on the choice of a plasmid, which should be able to allow the protein expression in the correct folding (similar to natural form) and requires an easy way to reproduce it (Routledge et al 2016). One of the alternatives for choosing the right plasmid for YSD is the construction of an expression cassette that has all the necessary components and allows the integration with conventional plasmids (Nasser et al 2003)
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