Bpm I

Enzyme name Bpm I
Prototype GsuI
Known analogs GsuI
SKU SE-E467
Turbo version Not available
High-concentration version Not available
Restriction type Type IIGiType IIG enzymes combine restriction and methyltransferase activities in a single polypeptide and are often stimulated by or dependent on AdoMet., Type 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'… CTGGAG(N)16▼ …3'
3'… GACCTC(N)14▲ …5'
Source Bacillus pumilus
Optimal buffer SE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
25507510050100
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; 50% glycerol. Store at -20°C
Ligation After 2-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and 95% may be recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity Blocked by overlapping Dcm-methylation (CmCWGG) 5'- C (5mC)TGG AG(N) 16 -3' 3'- G GA(5mC)C TC(N) 14 -5'
Reagents supplied with enzyme 10 X SE-buffer W, BSA
Notes To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  1. Unpublished data Degtyarev, S.Kh., Morgan P Unpublished observation (1992).
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