Roche Diagnostics Operations, Inc.

Boehringer Mannheim Gmbh

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Manfred Schmidt

Harald Sobek

Rainer Mueller

Johann-Peter Thalhofer

Bruno Frey

Michael Greif

Bernhard Suppmann

Markus Pajatsch

Christian Rudolph

Rainer Schmuck

Werner Hoelke

Klaus Kaluza

Bernd Schneidinger

Armin Ruf

Kirsten Sonn

Ingo Curdt

Peter Pallua

Brigitte Von Wilcken-Bergmann

Harlad Sobek

Daniel Schlieper

Benno Muller-Hill

The present disclosure provide novel variants of T7 RNA polymerase. Embodiments of T7 variants, according to the instant invention, include a Cysteine-Serine substitution on position 723 of the amino acid sequence of the T7 polypeptide. Embodiments of T7 variants according to the instant invention have a DNA-dependent RNA polymerase enzymatic activity and a reduced tendency to form intramolecular homodimers by way of oxidizing thiol groups. The amino acid substitutions within the T7 variants disclosed herein impact minimally, if at all, the RNA polymerase activity of the T7 polypeptide. Further, the mutations of the disclosed embodiments may optionally be combined with mutations which provide enhanced thermostability compared to the wild-type reference.

T7 RNA polymerase variants with cysteine-serine substitutions

The present invention provides improved variants of T7 RNA polymerase by introducing novel mutations which lead to improved thermostability of the enzyme. According to the invention, amino acid substitutions at the positions Val426, Ser633, Val650, Thr654, Ala702, Val795, and combinations thereof are advantageous.

T7 RNA polymerase variants with enhanced thermostability

The present invention provides an aqueous composition comprising a protein with enzymatic activity of alpha-galactosidase. The present invention further provides a method of stabilizing an aqueous composition comprising a protein with enzymatic activity of alpha-galactosidase, and a method of preparing a purified aqueous composition comprising the protein with enzymatic activity of alpha-galactosidase.

Stabilized aqueous alpha-galactosidase composition and methods relating thereto

Truncated terminal deoxynucleotidyl transferase (TdT) derivative from calf thymus, characterized in that the derivative in comparison to the native TdT is N-terminally truncated by up to 161 amino acids and has a 20- to 30-fold higher enzyme activity in solutions containing Coions, and its recombinant production and use.

Recombinant terminal deoxynucleotidyl transferase with improved functionality

The heterologous expression of the reverse transcriptase from the Avian Myeloblastosis Virus (AMV-RT) in prokaryotic cells and in particular() is described in the present invention. The invention also includes certain measures to simplify the purification of the heterodimeric AMV-RT.

Method for producing an active heterodimeric AMV-RT in prokaryotic cells

Thermolabile enzyme with uracil-DNA-glycosylase activity which is in particular characterized by a high degree of purity, short half-lives and a content of contaminating foreign activities of less than 2%, a process for its isolation as well as the use thereof to remove the base uracil from DNA and in particular from PCR products containing uracil. The enzyme is obtainable from gram-positive microorganisms such as e.g. Arthrobacter or Micrococcus.

Thermolabile uracil-DNA-glycosylas, process for its preparation and use for removing uracil from DNA

A vector according to the invention with a positive selection possibility for cloning a DNA sequence into a host cell transformed with the vector, wherein the vector contains suitable restriction cleavage sites within a selection gene the functional expression of which is prevented by insertion of the foreign gene is characterized in that it contains a modified cap gene as the selection gene which codes for a CAP protein with modified DNA binding specificity the expression of which is lethal for adenylate-cyclase-positive host cells.

Positive selection vector based on the caps gene, pCAPs vector and its

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Penzberg, Germany

Manfred Schmidt