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US 5,665,551 AGranted
Purified nucleic acid encoding a thermostable pyrophosphatase
AssigneeRoche Molecular Systems, Inc.
InventorsDavid Harrow Gelfand, Alice Ming Wang
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Abstract
Purified DNA sequences that encode a thermostable pyrophosphatase are provided. The purified DNA is obtained using a DNA probe consisting of SEQ ID NO: 1. Also provided are methods for producing thermostable pyrophosphatase.
Claims7
An isolated DNA consisting of SEQ ID NO:
- 1.
A method for isolating a target DNA fragment comprising a DNA coding for a thermostable pyrophosphatase from a bacterium of the genus Thermus, comprising:
- (a) forming a genomic library from said bacterium;
- (b) transforming or transfecting an appropriate host cell with the library of step (a);
- (c) contacting DNA from a transformed or transfected host cell of step
- (b) with a DNA probe consisting of SEQ ID NO:
- 1 under hybridization conditions consisting of the following:
- (1) hybridization:
- 6.times. SSPE, 2.times. Denhardt's reagent, 0.5% SDS, and 100 .mu.g/ml denatured, sheared salmon sperm DNA at 50.degree.-55.degree. C.;
- and
- (2) wash:
- 2.times. SSPE, 0.1% SDS for 15 minutes at 55.degree. C.;
- (d) isolating DNA which encodes a thermostable pyrophosphatase.
8 more elements
Dependencies are inferred from the claim text.