[Two site-specific endonucleases from the thermophilic OV Bacillus species strain]

Two site-specific endonucleases, BspOVI and BspOVII, were isolated from a thermophilic strain Bacillus species OV. The activities of both enzymes are maximal at 48 degrees C and do not depend on ATP and S-adenosyl-L-methionine. BspOVI recognizes the sequence [sequence: see text] and cleaves it as indicated by arrows. Thus, BspOVI is a IIN-subclass endonuclease isoshizomer of Eam1105I. BspOVI is very stable during storage. The enzyme can be used for direct T/A cloning of PCR products. BspOVII recognizes and cleaves the sequence [sequence: see text]; thus, BspOVII is an isoshizomer of CIaI. The cleavage by BspOVII is blocked by dam methylation of adenine inside the recognition site.