Thermostable nucleic acid polymerase from Thermococcus gorgonarius
AssigneeRoche Molecular Systems, Inc.
InventorsWaltraud Ankenbauer, Vitaly Svetlichny, Elizaveta Bonch-Osmolovskaya, Christine Ebenbichler, Bernhard Angerer, Gudrun Schmitz-Agheguian, Frank Laue
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Abstract
A purified thermostable enzyme is derived form the thermophilic archaebacterium Thermococcus gorgonarius. The enzyme can be native or recombinant, retains approximately 90% of its activity after incubation for two hours at 95° C. in the presence of stabilizing agents and possesses 3′-5′ proofreading exonuclease activity. Thermostable DNA polymerases are useful in many recombinant DNA techniques, especially nucleic acid amplification by the polymerase chain reaction (PCR).
Claims7
A process for preparation of a DNA polymerase, the process comprising:
- growing a microbial host cell comprising a DNA encoding a thermostable DNA polymerase obtainable from Thermococcus gorgonarius which catalyses the template directed polymerization of DNA, possesses 3′-5′-exonuclease (proofreading) activity and is characterized by at least a two-fold greater replication fidelity than DNA polymerase obtainable from Pyrococcus furiosus, wherein
- the DNA polymerase comprises the amino acid sequence set forth in SEQ ID NO:7;
- or
- the DNA comprises the nucleotide sequence set forth in SEQ ID NO:6;
- or
- the DNA polymerase has an apparent molecular weight between about 92,000 and 96,000 daltons;
- and
- purifying the DNA polymerase from the host cell.
Dependencies are inferred from the claim text.