What is FokI nuclease domain?
The nuclease consists of two separate domains, an N-terminal DNA-binding domain and a C-terminal DNA-cleavage domain. FokI exists as an inactive monomer in solution and becomes an active dimer upon binding to its target DNA and in the presence of specific divalent metals ( 11 ).
What is FokI enzyme?
FokI is a member an unusual class of restriction enzymes that recognize a specific DNA sequence and cleave nonspecifically a short distance away from that sequence. FokI consists of an N-terminal DNA recognition domain and a C-terminal cleavage domain.
How do you name an endonuclease?
Each enzyme is named after the bacterium from which it was isolated, using a naming system based on bacterial genus, species and strain. For example, the name of the EcoRI restriction enzyme was derived as shown in the box.
Is Fok1 a nuclease?
The restriction endonuclease Fok1, naturally found in Flavobacterium okeanokoites, is a bacterial type IIS restriction endonuclease consisting of an N-terminal DNA-binding domain and a non-specific DNA cleavage domain at the C-terminal.
What kind of enzyme is Fok in okeanokoites?
The enzyme Fok1 (Fok-1), naturally found in Flavobacterium okeanokoites, is a bacterial type IIS restriction endonuclease consisting of an N-terminal DNA-binding domain and a non-specific DNA cleavage domain at the C-terminal.
Where does the DNA binding domain of FokI come from?
Similar to ZFNs, TALENs are chimeric proteins composed of a DNA-binding domain fused to the nuclease domain of FokI. The DNA-binding domain used for TALENs is derived from transcription activator-like effectors (TALEs), which are naturally occurring proteins injected into host plant cells by Xanthomonas spp. bacteria.
Which is part of the DNA does FokI cleave?
FokI is a naturally occurring type IIS restriction enzyme and therefore cleaves outside of its DNA recognition sequence to one side. FokI has two distinct domains: a specific DNA-binding domain and a structurally separate DNA cleavage domain ( Kanazawa and Sugisaki, 1981 ).
Which is catalytic domain is required for FokI dimerization?
Fok I is a type IIs restriction endonuclease comprised of a DNA recognition domain and a catalytic domain. The structural similarity of the Fok I catalytic domain to the type II restriction endonuclease Bam HI monomer suggested that the Fok I catalytic domains may dimerize.