Izoschizomers

Fau I

Enzyme name

Fau I

Prototype

FauI

Known analogs

SmuI

SKU

SE-E209

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IISi

Recognition site

5'… CCCGC(N)₄^▼ …3'

3'… GGGCG(N)₆_▲ …5'

Source

An E.coli strain that carries the cloned Fau I gene from Flavobacterium aquatili

Optimal buffer

SE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)

Optimal temperature

55 °C

Inactivation temperature

65 °C

Buffer activity

B	G	O	W	Y	ROSE
100	25	0	0	50	50

Unit definition

One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°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% glycero; Store at -20°C.

Ligation

After 2-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and of these 95% can be recut.

Nonspecific hydrolysis

No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 55°C

Methylation sensitivity

Blocked by CG methylation.

Reagents supplied with enzyme

10 X SE-buffer B

Notes

The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,5.

References

V.A. Chernukhin, Y.G. Kashirina, K.S. Sukhanova, M.A. Abdurashitov, D.A. Gonchar, S.Kh. Degtyarev Isolation and exploration of biochemical propeties of DNA Methyltransferase FauIA modifying Non-palindromic sequence 5'-CCCGC-3'. // Biochemistry (Moscow), Vol. 70, No 6, (2005). (In Russian) 2005
M.A. Abdurashitov, S.S. Okhapkina, N.A. Netesova, L.N. Golikova, D.A. Gonchar, S.Kh. Degtyarev The unique FauI restriction-modification system: cloning and comparative analysis of protein structure // Molecular Biology (Moscow), Vol. 37, No.4, 619-624 (2003). (In Russian) 2003
S.Kh. Degtyarev, N.A. Netesova, V.E. Chizhikov, M.A. Abdurashitov Cloning and characterization of the gene encoding M.Fau I DNA methyltransferase. // Biol. Chem., Vol. 379, 567-568 (1998). 1998
V.S. Dedkov, S.Kh. Degtyarev, N.I. Rechkunova, V.E. Repin, V.A. Verkhozina, T.P. Vinogradova Flavobacterium acuatile - a strain producer of restriction endonuclease Fau I. // Soviet Union patent SU1661213 (1991). (In Russian) 1991
S.Kh. Degtyarev, A.A. Kolykhalov, N.I. Rechkunova, V.S. Dedkov Determination of substrate specificity of restriction endonuclease FauI. // Bioorg. chem (Moscow), Vol.15, No 1, 130-132 (1989). (In Russian) 1989