C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/55 (2006.01) C12N 1/21 (2006.01) C12N 9/16 (2006.01) C12N 9/22 (2006.01) C12N 15/62 (2006.01) C12N 15/70 (2006.01) C12Q 1/68 (2006.01)
Patent
CA 2170986
The present inventors have identified the recognition and cleavage domains of the FokI restriction endonuclease. Accordingly, the present invention relates to DNA segments encoding the recognition and cleavage domains of the FokI 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. One such hybrid restriction enzyme is Ubx-FN. This enzyme contains the homeo domain of Ubx linked to the cleavage or nuclease domain of FokI. Additionally, the present invention relates to the construction of two insertion mutants of FokI endonuclease.
Lesdits inventeurs ont identifié les domaines de reconnaissance et de clivage de l'endonuclease de restriction de FokI. Par conséquent l'invention concerne des segments d'ADN codant les domaines de reconnaissance et de clivage de l'endonucléase de restriction de FokI, respectivement. Le fragment N terminal de 41 kDa constitue le domaine de reconnaissance de FokI tandis que le fragment C-terminal de 25 kDa constitue le domaine nucléase de clivage de FokI. L'invention concerne également des enzymes de restriction hybride comprenant le domaine nucléase de l'endonucléase de restriction de FokI lié à un domaine de reconnaissance d'une autre enzyme. On peut citer comme enzyme de restriction hybride Ubx-F¿N?. Cette enzyme contient le domaine homéo de Ubx lié au domaine de clivage ou nucléase de FokI. De plus, l'invention concerne la construction de deux mutants d'insertion d'endonucléase de FokI.
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