HspA I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned HspA I gene from Haemophilus species A1
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
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by CG methylation 5′-G(5mC)GC-3’/3′-CG(5mC)G-5′.
Not blocked by methylation 5′-GCG(5mC)-3’/3′-CGCG-5′.
Cut hemimethylated site:5′- G(5mC)GC-3’/3′-CGCG-5`
Notes:
References:
Rechkunova, N.I., Prikhod’ko, E.A., Shevchenko, A.V., Degtyarev, S.K. Unpublished observations (1994).
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 4, pp 19-27, 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 // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.2, No 3, pp 39-46, 2006