| Autor: |
Tessier, Luc-Henri, Sondermeyer, Paul, Faure, Thérèse, Dreyer, Dominique, Benavente, Annie, Villeval, Dominique, Courtney, Michael, Lecocq, Jean-Pierre |
| Zdroj: |
Nucleic Acids Research; October 1984, Vol. 12 Issue: 20 p7663-7663, 1p |
| Abstrakt: |
Parameters influencing the efficiency of expression of the human ininune interferon (IFW-γ) gene in E.coli were studied by comparing a series of eight in vitro-derived gene variants. These contained all possible combinations of silent mutations in the first three codons of the mature IFN-γ polypeptide coding sequence. Expression levels varied up to 50-fold among the different constructions. Comparison of messenger RNA secondary structure models for each variant suggested that the presence of stem-loop structures blocking the translation initiation signals could drastically decrease the efficiency of IFN-γ synthesis. With variants displaying no stable n secondary structure in the region, a C+U transition at position +11 after the AUG resulted in a 5-fold increase in expression indicating that RNA primary structure also plays an important role in expression. In addition we demonstrate that, in this system, a spacing of 8 nucleotides between the Shine-Dalgarno region and AUG was optimal for gene expression and that the steady-state production level of IFN-γ rose exponentially with increasing rate of synthesis. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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