发明授权
- 专利标题: Method for screening restriction endonucleases
- 专利标题(中): 筛选限制性内切核酸酶的方法
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申请号: US09486356申请日: 2000-02-25
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公开(公告)号: US06383770B1公开(公告)日: 2002-05-07
- 发明人: Richard J. Roberts , Devon R. Byrd , Richard D. Morgan , Jay Patti , Christopher J. Noren
- 申请人: Richard J. Roberts , Devon R. Byrd , Richard D. Morgan , Jay Patti , Christopher J. Noren
- 主分类号: C12Q144
- IPC分类号: C12Q144
摘要:
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.
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