Izoschizomers

Fok I

Enzyme name

Fok I

Prototype

FokI

Known analogs

SKU

SE-E247

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IISi

Recognition site

5'… GGATG(N)₉^▼ …3'

3'… CCTAC(N)₁₃_▲ …5'

Source

An E.coli strain that carries the cloned Fok I gene from Flavobacterium okeanokoites

Optimal buffer

SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)

Optimal temperature

37 °C

Inactivation temperature

65 °C

Buffer activity

B	G	O	W	Y	ROSE
50	50	25	25	100	100

Unit definition

One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.

Assayed on

Lambda DNA

Storage conditions

10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.

Ligation

After 2-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.

Nonspecific hydrolysis

No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 1 u.a. of enzyme for 16 hours at 37°C

Methylation sensitivity

not tested

Reagents supplied with enzyme

10 X SE-buffer Y

Notes

Overdigestions of > 5 units of Fok I per 1μg of DNA and incubations > 2 hours are not recommended.

References

M.A. Abdurashitov, N.A. Netesova, L.N. Golikova, V.V. Gutorov, P.A. Belavin, S.Kh. Degtyarev The second DNA methyltransferase from BstF5I restriction-modification system is homologous to C-domains of methylases FokI and StsI. // Molecular Biology (Moscow), vol.33, p.87-94 (2000). (In Russian) 2000
L.N. Golikova, N.A. Netesova, V.V. Gutorov, P.A. Belavin, M.A. Abdurashitov, D.A. Gonchar, S.Kh. Degtyarev Multiplicity of site-specific DNA-methyltransferases of the BstF5I restriction modification system from Bacillus stearothermophilus F5. // Molecular Biology(Moscow), Vol.34, No.3, 443-447 (2000). (In Russian) 2000
M.A. Abdurashitov, E.V. Kileva, N.M. Shinkarenko, A.V. Shevchenko, V.S. Dedkov, S.Kh.Degtyarev BstF5 I, an unusual isoschizomer of Fok I. // Gene, 172, 4951 (1996). 1996
S.K. Degtyarev, P.A. Belavin, V.E. Repin, E.G. Malyvin Preparation method of restriction endonuclease Fok I from Flavobacterium okeanokoites // Soviet Union patent SU1436160 (1988). (In Russian) 1988
External publication Sugisaki, H., Kanazawa, S. Gene 16: 73-78 (1981). 1981