Expression of env sequences of the bovine leukemia virus (BLV) in the yeast Saccharomyces cerevisiae

Publisher Website
Google Scholar
Abstract
DNA sequences of the envelop (env) gene of the bovine leukemia virus (BLV) were expressed in the yeast saccharomyces cerevisiae. Two yeast promoters, the responsible PH05 promoter and the constitutive PGK promoter, were used to construct four expression plasmids either a sequence of the surface antigen gp51 or a (gp51 + gp30) sequence.The expressed hetrologous gene products were characterized by Western blot analysis and competitive radio‐immunoassay. By means of Northern blot analysis the steady‐state level of env‐specific mRNA was analysed.The highest expression rate was obtained from recombinant plasmid YEpSG 94 comprising a gp51 sequence—a 630 base pair fragment containing 70% of the gp51 but lacking the N terminus—as well as the PH05 promoter including PH05 signal sequence and the PH05 terminator. The recombinant gp51 was partially lycosylated but the PH05 signal peptide did not seem to be cleaved off. No immunoreactive material could be found in the periplasm or in the culture medium.By means of monoclonal antibodies directed against eight different epitopes of viral gp51, all for sequential antigenic determinants were detected in the AH 216(YEpSG 94) expression product.