Abstract
Production of a heat-stable, extracellular esterase by Streptomyces scabies is regulated by zinc ions. The esterase-encoding gene (est) from S. scabies was cloned and expressed in Streptomyces lividans. In S. lividans, expression of the est gene is also regulated by Zn2+, and the esterase is efficiently secreted in this organism. The sequence of the est gene suggests that a 39-amino acid signal peptide is removed during secretion of this protein. Deletion analysis has indicated that the hydrophobic domain of the signal peptide is required for secretion. Gel retardation assays and DNaseI footprinting using an S-30 protein extract from S. scabies have previously identified a specific 23-bp protein-binding site upstream from the est coding sequence. Deletion of this protein-binding sequence significantly decreased expression of the est gene.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 27-31 |
| Number of pages | 5 |
| Journal | Gene |
| Volume | 115 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Jun 15 1992 |
Bibliographical note
Funding Information:This work was supportedb y grantsf rom the National ScienceF oundation(D M~-8804638t)h, eH ermanF rasch Foundation(0 142-HF),a nd the AgriculturaEl xperiment Station at the Universityo f Minnesota.V .H. was supported by an NIH predoctoralt raininggrant (IT32-GM08347).
Keywords
- DNA-protein interaction
- Recombinant DNA
- gel retardation assay
- signal sequence
- zinc