Izoschizomers

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Sse9 I

Available supply options

Enzyme name

Sse9 I

Prototype

Tsp509I

Known analogs

MluCI, TasI, Tsp509I, TspEI

SKU

SE-E217

Turbo version

Available

High-concentration version

Not available

Restriction type

Type IIPi

Recognition site

5'… ^▼AATT …3'

3'… TTAA_▲ …5'

Source

An E.coli strain that carries the cloned Sse9 I gene from Sporosarcina species

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	75	50	50	75	75

Unit definition

One unit of the enzyme is the amount required to hydrolyze 1 μg of pBR322 DNA in 1 hour at 55°C in a total reaction volume of 50 μl.

Assayed on

pBR322 DNA

Storage conditions

10 mM Tis-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 5-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 DNA with 5 u.a. of enzyme for 16 hours at 55°C.

Methylation sensitivity

Blocked by methylation: 5`-A(m6A)TT-3`/ 3`-TT(m6A)A-5`.

Reagents supplied with enzyme

10 X SE-buffer B, BSA

Notes

At 37°C activity is 75% from maximum. To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.

References

D.A. Gonchar, M.A. Abdurashitov, S.S. Okhapkina, D.A. Shagin, E.V. Kileva, S.Kh. Degtyarev The Sse9I restriction-modification system: organization of genes and comparative analysis of proteins structures. // Molecular Biology, V.41, No 3, p. 491-498 (2007). (In Russian) 2007
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Ovchinnikov bulletin of biotechnology and physical and chemical biology V.3, No 4, p. 19-27.(2007) (In Russian) 2007
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
Y.P. Zernov, M.A. Abdurashitov, S.Kh. Degtyarev The Use of Restriction Analysis of Amplified 16S rRNA Genes for the Identification of Microorganisms: A Case Study of Bacterial Producers of Thermolabile Alkaline Phosphatase // Biotechnology, No. 6. – pp. 3–11 (2005). (In Russian) 2005
D.A. Gonchar, V.S. Dedkov, V.A. Verkhozina, Y.S. Kusner, A.V. Shevchenko, S.Kh. Degtyarev Sse9I, a Restriction Endonuclease from Sporosarcina sp. 9D, Recognizing the DNA Sequence 5′-AATT-3′. // Apllied Biochemistry and microbiology, Vol.34, No.2, 139-141 (1998). (In Russian) 1998
D.A. Gonchar, Yu. I. Wolf, S. Kh. Degtyarev Cloning and characterization of Sse9I DNA Methyltransferase recognizing 5′-AATT-3′. // Nucleic Acids Research, Vol. 24, No 14., 2790-2792 (1996). 1996