University of Georgia Research Foundation, Inc.

Joel Andrew Farkas

Janet Westpheling

Michael W W Adams

Daehwan Chung

Jennifer Huddleston

Gina L Lipscomb

The present invention relates to the discovery of a novel restriction/modification system in. The discovered restriction enzyme is a HaeIII-like restriction enzyme that possesses a thermophilic activity profile. The restriction/modification system also includes a methyltransferase, M.CbeI, that methylates at least one cytosine residue in the CbeI recognition sequence to mC. Thus, the invention provides, in various aspects, isolated CbeI or M.CbeI polypeptides, or biologically active fragments thereof; isolated polynucleotides that encode the CbeI or M.CbeI polypeptides or biologically active fragments thereof, including expression vectors that include such polynucleotide sequences; methods of digesting DNA using a CbeI polypeptide; methods of treating a DNA molecule using a M.CbeI polypeptide; and methods of transforming acell.

Restriction/modification polypeptides, polynucleotides, and methods

Provided herein are methods for transforming awith a polynucleotide. In one embodiment, the method includes contacting awith a polynucleotide under conditions suitable for uptake of the polynucleotide by the, and identifying transformants at a frequency of, for instance, at least 10transformants per microgram DNA. Also provided are isolatedhaving the characteristics ofCOM1, and plasmids that include an origin of replication that functions in a. The plasmid is stable in a recipientwithout selection for more than 100 generations and is structurally unchanged after replication infor more than 100 generations.

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Athens, GA, United States of America

Joel Andrew Farkas