Functional domains in flavobacterium okeanokoites (Foki) restriction endonuclease

摘要

The present inventors have identified the recognition and cleavage domains of the FokI restriction endonuclease. Accordingly, the present invention relates to the DNA segments encoding the recognition and cleavage domains of Fok I restriction endonuclease, respectively. The 41 kDa N-terminal fragment constitutes the FokI recognition domain while the 25 kDa C-terminal fragment constitutes the FokI cleavage nuclease domain. The present invention also relates to hybrid restriction enzymes comprising the nuclease domain of the FokI restriction endonuclease linked to a recognition domain of another enzyme. Additionally, the present invention relates to the construction of two insertion mutants of FokI endonuclease.