Merck Patent Gesellschaft MIT Beschrankter Haftung

Basf Aktiengesellschaft

Henkel Kommanditgesellschaft Auf Aktien

Gruenenthal Gmbh

Knoll Aktiengesellschaft

Tet Systems Holding Gmbh & Co. Kg

Wolfgang Hillen

Hermann Bujard

Manfred Gossen

Wolfgang Werner Strassburger

Gerd J Steffens

Hans G Gassen

Christiane Gatz

Stefanie Urlinger

Dirk Schnappinger

Vera Helbl

Ulrike Guelland

Robert Schmucker

Regina E Brigelius-Flohe'

Karl-Heinz Mauer

Martin R Wilhelm

Joachim Altschmied

Leopold Flohe'

The present invention provides a panel of transcriptional activator fusion proteins which comprises both tetracycline controlled transactivator proteins and reverse tetracycline transactivator proteins. These transactivators have novel phenotypes such as altered basal transcriptional activity in the absence of doxycycline, altered induced transcriptional activity in the presence of doxycycline, or differential induction by tetracycline and analogs of tetracycline.

Tet repressor-based transcriptional regulatory proteins

A &bgr;-Glucanase enzyme capable of hydrolytically cleaving mixed glucans is presented. The &bgr;-Glucanase is sufficiently stable under alkaline conditions for use in industrial cleaning processes, especially in the brewing industry.

&bgr;-glucanase from a bacillus

Plasmids containing synthetic DNA sequences are described which are suitable for the expression of the intermediate protein of the recombinant scu-PA (i.e., the unglycosylated protein moiety of the single chain prourokinase) in Enterobacteriaceae, especially in E. coli, with expression rates far higher than those obtainable according to prior methods. The plasmids comprise operons that include a regulatable, optionally synthetic, promotor, a Shine-Dalgarno sequence, a start codon, a synthetic structural gene with selected codon usage, and downstream of the structural gene, 1 to 2 transcription terminators. The preparation of the plasmids and of synthetic DNA-sequences starting from commercially available plasmids and intermediates is described. The intermediate protein, expressed in high yields by using the plasmids for transforming suitable hosts, may be refolded to produce the therapeutically useful plasminogen activator recombinant scu-PA. This protein also may serve as a starting material for obtaining the recombinant two chain unglycosylated urokinase protein (rtcu-PA) on a large scale by splitting the single chain product.

Plasmids, their construction and their use in the manufacture of a

Isolated nucleic acid molecules encoding fusion proteins which regulate transcription in eukaryotic cells are disclosed. The fusion proteins of the invention comprise a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells. Methods for regulating transcription of a tet operator-linked gene in a cell are also provided. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a fusion protein which regulates transcription, the fusion protein comprising a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells, and modulating the concentration of a tetracycline, or analogue thereof, in contact with the cell.

Tetracycline regulated transcriptional modulators with altered DNA

A polypeptide having mutarotase activity is obtained from a host microorganism that has been transformed with a molecule having a recombinant DNA sequence. The molecule having a recombinant DNA sequence is prepared by removing the first 60 nucleotides of a DNA sequence originating from the genome of Acinetobacter calcoaceticus that codes for the polypeptide, modifying the following 21 nucleotides and fusing of the resultant structural gene with the start of the tetracycline-repressor gene and with an effective promotor sequence. The tetracycline-regressor gene and the promotor sequence preferably originate from the same microorganism such as E. coli in which expression of the polypeptide is carried out, and result in increased yield of the expressed polypeptide having mutarotase activity.

Recombinant DNA, transformed host microorganism and method for producing

The invention relates to DNA sequences, recombinant DNA molecules and transformed host organisms, and to their use in a process for the genetic engineering preparation of a polypeptide having the biological activity of the enzyme mutarotase.

DNA sequences, recombinant DNA molecules and process for the preparation

Growing community of inventors

Erlangen, Germany

Wolfgang Hillen