AsuHP I

Enzyme nameAsuHP I
PrototypeHphI
Known analogsHphI
SKUSE-E231
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GGTGA(N)8▼ …3'
3'… CCACT(N)7▲ …5'
SourceAn E.coli strain that carries the cloned AsuHPI gene from Actinobacillus suis HP
Optimal bufferSE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10-2550-7510075-10025-50100
Unit definitionOne unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.
Assayed onLambda DNA (dam-)
Storage conditions10 mM Tris-HCl (pH 7.5), 250 mM NaCl, 0.1 mM EDTA, 7 mM 2-mercaptoethanol, 200 µg/ml BSA, 50% glycerol; Store at -20°С.
LigationAfter 5-fold overdigestion with AsuHP I, ~30% of the DNA fragments can be ligated with T4 DNA Ligase and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by overlapping dam-methylation (GmATC) GGT GA TC
Reagents supplied with enzyme10 x SE-Buffer O
Notesenzyme may cleave at N9/N8 depending on the sequence between the ecognition and cleave sites.
References
  2. V.S. Dedkov, S.Kh. Degtyarev Actinobacillus and Streptococcus: producers of isoschizomers of the restriction endonucleases R.Hph I, R.Sau I, R.Nhe I, R.Mbo I and R.Swa I. // Biol. Chem., Vol. 379, 573-574, (1998).
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