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Nucleic Acids Research, 1985, Vol. 13, No. 20 7171-7182
© 1985

Articles

Purification of Mbo II methylase (GAAGmA from Moraxella bovis: site specific cleavage of DNA at nine and ten base pair sequences

Michael McClelland, Michael Nelson* and Charles R. Cantor

Department of Human Genetics and Development, Columbia University 701 W. 168th St., New York, NY 10032 *New England Biolabs 32 Tozer Rd., Beverly, MA 01915, USA

Received September 20, 1985. Accepted September 26, 1985.

The restriction modification methylase M.Mbo II has been purified using a sensitive oligonucleotide linker assay. The enzyme methylates the Mbo II recognition sequence GAAGA at adenine to produce GAAGmA. M. Mbo II can be used in conjunction with the methylation dependent restriction endonuclease Dpn I (GmATC) to produce cleavage at the 10 base sequence GAAGATCTTC. When M.Mbo II is used in combination with M.Cla I (ATCGATCGAT), cleavage by Dpn I occurs at the four ten base sequences GAAGATCTTC, GAAGATCGAT, ATCGATCTTC and ATCGATCGAT, which is equivalent to a nine base recognition site. The use of combinations of adenine methylases and Dpn I to generate highly selective DNA cleavages at a variety of sequences up to fourteen base pairs is discussed.

* For all sequences only one strand is shown, oriented 5' to 3'. mA represents 6-methyladenine.


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