| Title: | Efficient synthesis of secreted murine interleukin-2 by Saccharomyces cerevisiae: influence of 3'-untranslated regions and codon usage |
| Author(s): | Demolder J; Fiers W; Contreras R; |
| Address: | "Laboratory of Molecular Biology, State University, Gent, Belgium" |
| DOI: | 10.1016/0378-1119(92)90688-l |
| ISSN/ISBN: | 0378-1119 (Print) 0378-1119 (Linking) |
| Abstract: | Several expression vectors were compared which directed the synthesis of secreted murine interleukin-2 (mIL2) in the culture medium of Saccharomyces cerevisiae. We used the prepro-sequence of the alpha 1 mating-factor precursor as a secretion signal in S. cerevisiae in combination with different promoters. The yield of mature mIL2 was significantly improved by deleting the major part of the 3'-untranslated region (UTR). In Northern-blotting experiments we showed that a destabilizing sequence present in the 3' UTR might be responsible for rapid degradation of the mIL2 mRNA. The highest expression (about 10 micrograms/ml) was obtained under control of the GAL1 promoter in an S. cerevisiae strain where the regulatory GAL4 gene was overexpressed. No difference in expression level was observed in a construct wherein twelve consecutive codons were replaced by optimal codons for S. cerevisiae |
| Keywords: | "Animals Base Sequence Blotting, Northern Codon/*genetics DNA-Binding Proteins Fungal Proteins/genetics Gene Expression Regulation, Fungal/*genetics Genetic Vectors/*genetics Interleukin-2/biosynthesis/*genetics/metabolism Mating Factor Mice Molecular Sequ;" |
| Notes: | "MedlineDemolder, J Fiers, W Contreras, R eng Research Support, Non-U.S. Gov't Netherlands 1992/02/15 Gene. 1992 Feb 15; 111(2):207-13. doi: 10.1016/0378-1119(92)90688-l" |
