| 摘要 |
The invention provides improvements in alpha-factor leader peptide-facilitated secretion of polypeptides from S. cerevisiae into culture media. Recombinant expression vectors suitable for transformation of S. cerevisiae are provided which encode a fusion polypeptide including the pre-pro-alpha-factor leader peptide, or modification thereof, and the heterologous polypeptide to be secreted. Culture of S. cerevisiae strains transformed with an expression vector of the invention leads to secretion to the culture medium of heterologous polypeptide of interest. Increased recoveries of certain polypeptides, such as GRF, which retain their biological activities, are obtained by modifying peptide-encoding DNA sequences on the expression vector to produce analog polypeptides which are less subject proteolytic degradation or other activity-reducing modifications during and after secretion. a-strains of S. cerevisiae are provided which, upon transformation with an expression vector according to the invention, with a fused gene controlled by the alpha-factor promoter, followed by culturing, secrete heterologous polypeptide in an alpha-factor-mediated secretion process. Peptide recovery is also enhanced by employing as host for transformation a S. cerevisiae strain with impaired proteolytic function. |
