Thermostable polymerases having altered fidelity and method of identifying and using same
US6395524B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Nov 26, 1997 |
| Grant date | May 28, 2002 |
| Priority date | — |
| Expiry date | Nov 26, 2017 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6883
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides a method for identifying a thermostable polymerase having altered fidelity. The method consists of generating a random population of polymerase mutants by mutating at least one amino acid residue of a thermostable polymerase and screening the population for one or more active polymerase mutants by genetic selection. For example, the invention provides a method for identifying a thermostable polymerase having altered fidelity by mutating at least one amino acid residue in an active site O-helix of a thermostable polymerase. The invention also provides thermostable polymerases and nucleic acids encoding thermostable polymerases having altered fidelity, for example, high fidelity polymerases and low fidelity polymerases. The invention additionally provides a method for identifying one or more mutations in a gene by amplifying the gene with a high fidelity polymerase. The invention further provides a method for accurately copying repetitive nucleotide sequences using a high fidelity polymerase mutant. The invention also provides a method for diagnosing a genetic disease using a high fidelity polymerase mutant. The invention further provides a method for r…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.