High level of expression of a protective antigen of schistosomes in Saccharomyces cerevisiae.
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.
Loison
G
G
Transgene S.A., Strasbourg, France.
Vidal
A
A
Findeli
A
A
Roitsch
C
C
Balloul
J M
JM
Lemoine
Y
Y
D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000302:isolation & purification / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012550:Schistosoma mansoni / Q000276:immunology*; D014170:Transformation, Genetic
D000947:Antigens, Helminth; D011994:Recombinant Proteins; D005982:Glutathione Transferase