High level of expression of a protective antigen of schistosomes in Saccharomyces cerevisiae

Yeast. 1989 Nov-Dec;5(6):497-507. doi: 10.1002/yea.320050609.

Abstract

Strains of Saccharomyces cerevisiae expressing P28-I, an antigen inducing protection against schistosomiasis, have been constructed. Transformants containing a very high copy number of a P28-I expression vector were selected by genetic complementation involving deficient LEU2 or URA3 alleles carried by plasmids. Using the ura3 fur1 auto-selection system, constitutive and stable expression of P28-I could be obtained in cultures grown in rich medium. The accumulation of the foreign protein exceeds 25% of total yeast proteins when estimated by Coomassie Brilliant Blue staining of SDS-PAGE. Moreover, P28-I which was located intracellularly was soluble and biologically active.

MeSH terms

  • Animals
  • Antigens, Helminth / biosynthesis*
  • Antigens, Helminth / genetics
  • Base Sequence
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Glutathione Transferase / isolation & purification
  • Glutathione Transferase / metabolism
  • Plasmids
  • Recombinant Proteins / biosynthesis
  • Saccharomyces cerevisiae / metabolism*
  • Schistosoma mansoni / immunology*
  • Transformation, Genetic

Substances

  • Antigens, Helminth
  • Recombinant Proteins
  • Glutathione Transferase