Izoschizomers

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Kzo9 I

Available supply options

Enzyme name

Kzo9 I

Prototype

MboI

Known analogs

BfuCI, Bsp143I, BstMB I, DpnII, MboI, NdeII, Sau3AI, BstKT I

SKU

SE-E187

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IIPi

Recognition site

5'… ^▼GATC …3'

3'… CTAG_▲ …5'

Source

Kurthia zopfii 9

Optimal buffer

SE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)

Optimal temperature

37 °C

Inactivation temperature

65 °C

Buffer activity

B	G	O	W	Y	ROSE
50	100	50	50	50	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 KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol; Store at -20°C.

Ligation

After 3-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 6 u.a. of enzyme for 16 hours at 37°C.

Methylation sensitivity

Not blocked by overlapping Dam methylation (G m ATC) GATC. Blocked by overlapping CG methylation C G AT m C G. Cleaved of DNA is impaired by overlapping CG methylation GAT m C G.

Reagents supplied with enzyme

10 X SE-buffer G

References

A.N. Nadeev, V.A. Chernukhin, O.O. Sevastyanova, Y.E. Tomilova, N.M. Shinkarenko, A.A. Evdokimov, S.Kh. Degtyarev BstKTI, a New dam-Sensitive Neoschizomer of Restriction Endonuclease MboI, which is Able to Cleave Hemimethylated Substrate. // Biotekhnologia (Moscow), No.2, 5-10 (2006). (In Russian) 2006
M.A. Abdurashitov, V.N. Tomilov, V.A. Chernukhin, D.A. Gonchar, S.Kh. Degtyarev Mammalian chromosomal DNA digestion with restriction endonucleases in silico // Ovchinnikov bulletin of biotechnology and physical and chemical biology , V.2, No 3, p. 29-38 (2006). (In Russian) 2006
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // Ovchinnikov bulletin of biotechnology and physical and chemical biology , V.2, No 3, p. 39-46 (2006). (In Russian) 2006
S.Kh. Degtyarev, N.I. Rechkunova, A.A. Grinev, V.S. Dedkov Discovery and substrate specificity determination of restriction endonucleases Bme18I and Kzo9I. // Izvestia SB SA USSR, Biological sciences series, No.3, 25-26 (1989). (In Russian) 1989
S.Kh. Degtyarev, N.I. Rechkunova, O.A. Zhilkina, G.V. Semenchenko, M.I. Novozhilova Kurthia zopfii - a strain producer of restriction endonuclease Kzo9I // Soviet Union patent SU 1440919 (1988). (In Russian) 1988