Engineering nicking endonucleases from type IIs restriction endonucleases
US6395523B1 · kind B1 · utility
16Cited by
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7Claims
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Key dates
| Filing date | Jun 1, 2001 |
| Grant date | May 28, 2002 |
| Priority date | — |
| Expiry date | Jun 1, 2021 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N9/22
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention relates to methods to engineer nicking endonucleases from existing Type IIs restriction endonucleases, and the production of the engineered nicking endonucleases. Two engineering methods are disclosed. One involves inactivating the dimerization function of a Type IIs restriction enzyme using site-directed mutagenesis approach. The other involves replacing the cleavage domain of a Type IIs restriction enzyme with the cleavage domain from a natural occurring nicking endonuclease, N.BstNBI.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.