Roche Diagnostics Operations, Inc.

Thomas Kirschbaum

Rainer Mueller

Johann-Peter Thalhofer

Werner Hoelke

Frank Geipel

Stephan Glaser

Walter Eberle

Bettina Bommarius

Hellmut Eckstein

Helmut Schoen

Michael Knoll

The present invention deals with a method of preparing nucleic acids, particularly RNA, from a whole blood sample. The nucleic acids purified by the method of the invention are particularly suited for detection of nucleic acid marker molecules. Preferred are markers for the diagnosis of transmissible spongiform encephalopathy (TSE). Such diagnosis is based on the detection, by means of real-time PCR, of certain mRNAs of the TSE-infected organism. Said mRNAs specifically originate as splicing variants and are isolated from whole blood by the method of the invention.

Nucleic acid preparation from whole blood for use in diagnosis of transmissible spongiform encephalopathy

The invention concerns a method for the expression of a gene coding for a soluble proteinase K in yeast e.g. inwith subsequent secretion into the culture medium. In addition a method for purifying the heterologously expressed and secreted proteinase K is described.

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The invention concerns a method for the recombinant production or expression of eukaryotic alkaline phosphatase mutants in yeast cells wherein the specifically introduced mutations result in a reduction of the specific AP activity by at least a factor 1:100. The invention also concerns a method for inserting corresponding nucleic acid sequences into a vector for expression in methylotrophic yeast strains and it concerns corresponding vectors and host strains.

Production of weakly active or inactive mutants of alkaline phosphatase and their expression in yeast

This invention involves a process for selecting and producing eukaryotic alkaline phosphatase in yeast. Yeast cells are subjected to multiple transformations using a vector comprising a first resistance marker gene and the alkaline phosphatase gene. Those strains that grow in media containing the first resistance marker are further transformed using a vector comprising a second selection marker gene and the alkaline phosphatase gene. Transformants that grow in media containing the second selection marker are selected for expressing the eukaryotic alkaline phosphatase.

Expression of alkaline phosphatase in yeast

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Munich, Germany

Thomas Kirschbaum