Factors affecting heterologous gene expression in Saccharomyces cerevisiae.
The 'promoter' fragment from the yeast phosphoglycerate kinase (PGK) gene has been used to direct the expression of human interferon-alpha-2 (IFN alpha 2) on a high-copy-number plasmid in yeast. The yields of IFN alpha 2 are only 1-3% of yeast total protein, whereas the maximum yield of PGK produced by the PGK gene on a high-copy-number plasmid is at least 50%. IFN alpha 2 is turned over more rapidly than PGK but in addition a major reason for the relatively low level of IFN alpha 2 is that IFN-specific RNA levels are much lower. This does not reflect differences in plasmid copy number or integrity, or differences in the 5' and 3' untranslated regions of the transcripts or DNA flanking regions. It appears that the presence of heterologous coding sequences, or the absence of specific yeast sequences causes a reduction in heterologous RNA levels in yeast.
D003001:Cloning, Molecular; D004261:DNA Replication; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D007370:Interferon Type I / Q000235:genetics*; D010735:Phosphoglycerate Kinase / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004274:DNA, Recombinant; D007370:Interferon Type I; D012333:RNA, Messenger; D010735:Phosphoglycerate Kinase