Method for screening restriction endonucleases
US6383770B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Feb 25, 2000 |
| Grant date | May 7, 2002 |
| Priority date | — |
| Expiry date | Feb 25, 2020 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/922
- WIPO fieldMachine tools
- WIPO sectorMechanical engineering
Abstract
A method is provided for identifying a restriction endonuclease, which includes the steps of (a) screening a target DNA sequence for the presence of known methylase sequence motifs, (b) identifying any open reading frames which lie close to the methylase sequence motifs screened in step (a), and (c) assaying the protein products of these open reading frames for restriction endonuclease activity. Methods for identifying isoschizomers of known restriction endonucleases, which isoschizomers possess a desired physical property, such as thermostability, are also provided by the present invention, as are several novel restriction endonucleases isolated from M. jannaschii, MjaIII and MjaIV. Additionally, a gene was identified that encoded a previously observed endonuclease activity, designated MjaII. Also provided by the present invention are vectors suitable for cloning a DNA sequence encoding a cytotoxic protein, via independent transcription promotors which may be selectively controlled by several conditions. A method for producing these cytotoxic proteins using such vectors is also provided, as are stable clones of PacI and NlaIII.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.