Patent ID: 6083723
Filing Date: 2000-07-04
Classification: A61K,A61L,A61P,C07K,C12N

Abstract:
A vector comprising a nucleotide sequence that comprises in the 5' to 3' direction and operably linked (a) a yeast-recognized transcription and translation initiation region, (b) a coding sequence for a hybrid precursor polypeptide, and (c) a yeast-recognized transcription and translation termination region, wherein said hybrid precursor polypeptide comprises:5'-SP-(PS).sub.n-1 -(LP-PS).sub.n-2 -(NPRO.sub.MHP -PS).sub.n-3 -MHP-(PS-CPRO.sub.MHP).sub.n-4 -3'wherein:SP comprises a signal peptide sequence for a yeast secreted protein;PS comprises a preferred processing site cleaved in vivo by a yeast proteolytic enzyme;LP comprises a leader peptide sequence for a yeast secreted protein;NPRO.sub.MHP comprises a native N-terminal propeptide sequence of a mature heterologous mammalian protein of interest;MHP comprises a peptide sequence for said mature heterologous mammalian protein of interest;CPRO.sub.MHP comprises a native C-terminal propeptide sequence of said mature heterologous mammalian protein of interest; andn-1, n-2, n-3, and n-4 independently=0 or 1;wherein said mammalian protein is a variant PDGF-BB protein, said variant having an amino acid sequence that has at least about 90% sequence identity to the amino acid sequence of a native PDGF-BB protein, wherein said processing sites allow for proteolytic processing of said precursor polypeptide to said mature protein in vivo by a yeast host cell, and wherein at least n-3 or n-4=1.