Izoschizomers

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BstF5 I

Available supply options

Enzyme name

BstF5 I

Prototype

(FokI)

Known analogs

BseGI, Fok I

SKU

SE-E031

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IISi

Recognition site

5'… GGATGNN^▼ …3'

3'… CCTAC_▲NN …5'

Source

Bacillus stearothermophilus F5

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

65 °C

Inactivation temperature

80 °C

Buffer activity

B	G	O	W	Y	ROSE
75	50	25	50	100	100

Unit definition

One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°C in a total reaction volume of 50 μl.

Assayed on

Lambda DNA

Storage conditions

10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol; Store at -20°C.

Ligation

After 10-fold overdigestion with enzyme more than 90% 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 20 u.a. of enzyme for 16 hours at 65°C.

Methylation sensitivity

not tested

Reagents supplied with enzyme

10 X SE-buffer Y

Notes

The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,25.

References

V.A. Chernukhin, V.V. Kuznetsov, D.A. Gonchar, Y.G. Kashirina, N.A. Netesova, S.Kh. Degtyarev Analysis of substrate specificity and biochemical properties of DNA methyltransferase M3.BstF5I from BstF5I restriction-modification system // Biochemistry (Moscow), Vol. 75, No 1, p.78-87 (2010). (In Russian) 2010
V.A. Chernukhin, L.N. Golikova, D.A. Gonchar, M.A. Abdurashitov, Y.G. Kashirina, N.A. Netesova, S.Kh. Degtyarev M.BstF5I-2 and M.BstF5I-4 DNA Methyltransferases from BstF5I restriction-modification system of Bacillus stearothermophilus F5. // Biochemistry (Moscow), Vol. 68, No 9, (2003). (In Russian) 2003
N.A. Netsova, L.N. Golikova, L.G. Ovechkina, A.A. Evdokimov, E.G. Malygin, N.S. Gololobova, D.A. Gonchar, S.Kh. Degtyarev The restriction-modification system from Bacillus stearothermophilius F5: Comparison study of the DNA methyltransferases M.BstF5I-1 and M.BstF5I-3. // Molecular Biology(Moscow), vol.36, p.1-8 (2002). (In Russian) 2002
L.V. Golikova, V.V. Gutorov, A.A. Evdokimov, S.N. Shchelkunov, D.A. Gonchar, S.S. Okhapkina, S.Kh. Degtyarev, N.A. Netesova M.BstF5I-1, the forth DNA-methyltransferase of the BstF5I restriction-modification system from Bacillus stearothermophilius F5. // Biorg. Chemistry(Moscow), vol.28, p.84-86 (2002). (In Russian) 2002
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
S.S. Okhapkina, N.A. Netesova, L.V. Golikova, E.V. Seregina, S.V. Sosnovtsev, M.A. Abdurashitov, 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
S.Kh. Degtyarev, N.A. Netesova, M.A. Abdurashito, A.V. Shevchenko Primary structure and strand specificity of BstF5 I — 1 DNA methyltransferase which recognizes 5′-GGATG-3′ // Gene, 187, 217-219 (1997). 1997
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