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Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae.
Piraine, Renan Eugênio Araujo; Gonçalves, Vitória Sequeira; Dos Santos Junior, Alceu Gonçalves; Cunha, Rodrigo Casquero; de Albuquerque, Pedro Machado Medeiros; Conrad, Neida Lucia; Leite, Fábio Pereira Leivas.
Affiliation
  • Piraine REA; Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
  • Gonçalves VS; Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
  • Dos Santos Junior AG; Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
  • Cunha RC; Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
  • de Albuquerque PMM; Programa de Pós-Graduação em Veterinária, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
  • Conrad NL; Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
  • Leite FPL; Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Biotechnol Lett ; 43(8): 1649-1657, 2021 Aug.
Article in En | MEDLINE | ID: mdl-33934257
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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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Plasmids / Saccharomyces cerevisiae / Recombinant Fusion Proteins / Cell Surface Display Techniques Limits: Animals Language: En Journal: Biotechnol Lett Year: 2021 Document type: Article Affiliation country: Brazil Country of publication: Netherlands

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Plasmids / Saccharomyces cerevisiae / Recombinant Fusion Proteins / Cell Surface Display Techniques Limits: Animals Language: En Journal: Biotechnol Lett Year: 2021 Document type: Article Affiliation country: Brazil Country of publication: Netherlands