- 专利标题: Thermostable polymerases having altered fidelity and methods of identifying and using same
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申请号: US12939798申请日: 2010-11-04
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公开(公告)号: US08420325B2公开(公告)日: 2013-04-16
- 发明人: Lawrence A. Loeb , Leroy Hood , Motoshi Suzuki
- 申请人: Lawrence A. Loeb , Leroy Hood , Motoshi Suzuki
- 申请人地址: US WA Seattle
- 专利权人: University of Washington Through Its Center for Commercialization
- 当前专利权人: University of Washington Through Its Center for Commercialization
- 当前专利权人地址: US WA Seattle
- 代理机构: Nixon Peabody LLP
- 代理商 Mark J. FitzGerald
- 主分类号: C12Q1/68
- IPC分类号: C12Q1/68 ; C12P19/34 ; C07K1/00
摘要:
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 randomly mutagenizing a gene by amplifying the gene using a low fidelity polymerase mutant.
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