Izoschizomers

Psp6 I

Enzyme name

Psp6 I

Prototype

EcoRII

Known analogs

Ajn I, EcoRII, PspGI, BptI, BseBI, Bst2U I, BstNI, BstOI, MvaI

SKU

SE-E453

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IIPi

Recognition site

5'… ^▼CCWGG …3'

3'… GGWCC_▲ …5'

Source

Pseudomonas species 6

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

80 °C

Buffer activity

B	G	O	W	Y	ROSE
100	50	10	25	75	10

Unit definition

One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-) in 1 hour at 55°C in a total reaction volume of 50 μl.

Assayed on

Lambda DNA (dcm-)

Storage conditions

10 mM Tris-HCl (pH 7.5); 100 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 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 4 u.a. of enzyme for 16 hours at 37°C.

Methylation sensitivity

Blocked by overlapping Dcm methylation(C m CWGG) CCWGG.

Reagents supplied with enzyme

10 X SE-buffer B

Notes

At 37°C, the activity is very low.

References

V.S. Dedkov, N.A. Mikhnenkova, L.P. Vlasenko, Y.E. Tomilova, Y.G. Kashirina, D.A. Gonchar, S.Kh. Degtyarev Restriction Endonuclease AjnI from Acinetobacter johnsonii R2, an Isoschizomer of EcoRII, recognizes 5′- ↓CCWGG-3′ and cleaves dcm-methylated sites. // Biotekhnologia (Moscow), No.3, 19-24 (2004). (In Russian) 2004
Unpublished data Dedkov, V.S., Tomilova, J.E., Kileva, E.V., Degtyarev, S.K Unpublished observations. (2002) 2002