Tetracycline promoter for the stringently regulated production of recombinant proteins in prokaryotic cells
AssigneeMax-Planck-Gesellschaft zur Forderung der Wissenschaften e.V.
InventorsArne Skerra, Christina Wardenberg
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Abstract
The present invention concerns a prokaryotic vector which contains a regulatable expression control sequence that can be repressed by the repressor of the tetracycline resistance gene, a prokaryotic cell transformed with this vector and the use of the vector or the cell in a process for the production of polypeptides in prokaryotes by genetic engineering.
Claims25
A prokaryotic vector comprising (a) a regulatable expression control sequence which can be repressed by a tetracycline repressor protein and (b) a DNA encoding a tetracycline repressor protein in operative linkage with an expression control sequence which cannot be repressed by a tetracycline repressor protein.
The plasmid pASK75 having the nucleotide sequence shown in SEQ ID NO.2.
A prokaryotic cell containing (a) a gene coding for a heterologous polypeptide in operative linkage with a regulatable expression control sequence which can be repressed by a tetracycline repressor protein and (b) a DNA encoding a tetracycline repressor protein in operative linkage with an expression control sequence which cannot be repressed by a tetracycline repressor protein, wherein said cell is transformed with at least one copy of a vector comprising (a) a DNA encoding a regulatable expression control sequence which can be repressed by a tetracycline repressor protein and (b) a tetracycline repressor protein in operative linkage with an expression control sequence which cannot be repressed by a tetracycline repressor protein.
A process for the production of polypeptides in a prokaryotic cell by genetic engineering, comprising the steps of:
- transforming a cell with (a) at least one gene coding for a heterologous polypeptide in operative linkage with a regulatable expression control sequence that can be repressed by a tetracycline repressor protein and (b) a DNA encoding a tetracycline repressor protein in operative linkage with an expression control sequence that cannot be repressed by a tetracycline repressor protein,
- culturing said cell under conditions which lead to an expression of the gene coding for the heterologous peptide, and
- isolating the heterologous polypeptide from the cell or from the medium.
Dependencies are inferred from the claim text.