BmoI Restriction Endonuclease from the Bacterial Strain Brevundimonas mongoliensis 53 Recognizes the DNA Sequence 5’-GG^GWCCC-3’
We identified a bacterial strain, Brevundimonas mongoliensis 53, which produces a new Type II restriction endonuclease, designated BmoI.
BmoI recognizes the palindromic heptanucleotide sequence 5’-GG^GWCCC-3’ (W = A or T) and cleaves at the indicated position to generate three-nucleotide 5′ overhangs.
BmoI is an isoschizomer of the previously described restriction enzyme SanDI.
The producing strain was identified based on morphological and biochemical characteristics and by sequencing a fragment of the 16S rRNA gene.
A preparation of BmoI with an activity of 2000 U/ml was obtained using a three-step chromatographic purification workflow.
Optimal reaction conditions for BmoI were determined to be SE buffer W (10 mM Tris-HCl, pH 8.5; 10 mM MgCl₂; 100 mM NaCl; 1 mM DTT) and an incubation temperature of 37°C.