发明名称 |
Method for engineering strand-specific, sequence-specific, DNA-nicking enzymes |
摘要 |
Methods are provided for converting into a sequence specific strand specific and location specific DNA nicking endonuclease, a restriction endonuclease that recognizes an asymmetric DNA sequence, the endonuclease having two catalytic sites and one or more single sequence specific DNA-binding domains. In one embodiment the method requires inactivating one of the catalytic sites of the restriction endonuclease. In another embodiment, the restriction endonuclease is a dimer having a first and second subunit each comprising a sequence specific DNA binding domain, a catalytic site and a dimerization domain. The nicking endonuclease is formed from combining one subunit having an inactivated catalytic site and a second subunit having an inactivated DNA binding domain. The nicking endonuclease may be converted into a restriction endonuclease by the addition of manganese cations in the digestion buffer.
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申请公布号 |
US7081358(B2) |
申请公布日期 |
2006.07.25 |
申请号 |
US20020223074 |
申请日期 |
2002.08.16 |
申请人 |
NEW ENGLAND BIOLABS, INC. |
发明人 |
HEITER DANIEL;LUNNEN KEITH;WILSON GEOFFREY G. |
分类号 |
C12N15/55;C12N9/22;C12P21/04 |
主分类号 |
C12N15/55 |
代理机构 |
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