Recombinant bovine pancreatic desoxyribonuclease I with high specific activity
AssigneeRoche Diagnostics Operations, Inc.
InventorsBernhard Suppmann, Johann-Peter Thalhofer, Stephanie Ronning, Helmut Schoen, Rainer Müller
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Abstract
A method is provided for the production of a bovine pancreatic protein with a specific desoxyribonuclease activity of at least 6,000 units per mg of protein. Methylotrophic yeast is used as a heterologous host organism. The bovine pancreatic protein is secreted into the growth medium from which it is purified.
Claims19
A method of producing a bovine pancreatic protein with a specific desoxyribonuclease activity of at least 6,000 units per mg of protein purified in step (d), comprising the steps of
- (a) providing a vector comprising a nucleotide sequence that encodes a pre-protein, the pre-protein comprising the bovine pancreatic protein and a signal peptide,
- (b) transforming a methylotrophic yeast strain with the vector,
- (c) cultivating the transformed methylotrophic yeast strain in a growth medium that comprises nutrients and methanol, whereby the methylotrophic yeast strain expresses and secretes the bovine pancreatic protein into the growth medium, and
- d) purifying the bovine pancreatic protein from the growth medium.
A kit containing a bovine pancreatic protein with a specific desoxyribonuclease activity of at least 6,000 units per mg of protein and a reaction buffer comprising a divalent cation.
A method of producing a bovine pancreatic protein comprising the steps of:
- (a) providing a vector comprising a nucleotide sequence comprising the sequence of SEQ ID NO:
- 6 operably linked to the sequence of SEQ ID NO:
- 5 that encodes the bovine pancreatic protein of SEQ ID NO:
- 2,
- (b) transforming a methylotrophic yeast strain with the vector,
- (c) cultivating the transformed methylotrophic yeast strain in a growth medium that comprises nutrients and methanol whereby the methylotrophic yeast strain expresses and secretes the bovine pancreatic protein into the growth medium, and
- (d) purifying the bovine pancreatic protein from the growth medium.
Dependencies are inferred from the claim text.