Tetracycline-inducible transcriptional activator and tetracycline-regulated transcription units

US5654168A · kind A · utility

Assignees

• BASF SE
• Knoll, Inc. · DE

Inventors

• Hermann Bujard · Heidelberg, DE
• Manfred Gossen · Corona, US

Key dates

• Filing date: Jul 15, 1994
• Grant date: Aug 5, 1997
• Expiry date: Jul 15, 2014

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2830/006
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Nucleic acid molecules and proteins useful for regulating the expression of genes in eukaryotic cells and organisms in an inducible manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is stimulated by a transcriptional activator fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences in the presence of tetracycline or a tetracycline analogue and a second polypeptide which directly or indirectly activates transcription in eukaryotic cells. In one embodiment, the fusion protein comprises a mutated Tet repressor operatively linked to a transcriptional activation polypeptide, such as a portion of herpes simplex virus virion protein 16. In the absence of an inducing agent (tetracycline or a tetracycline analogue), transcription of the tet operator-linked nucleotide sequence remains uninduced. In the presence of the inducing agent, transcription of the tet operator-linked nucleotide sequence is stimulated by the transactivator fusion protein of the invention. Novel transcription units which allow for coordinate or independent tetracycline-regulated expression of two or m…