New England Biolabs, Inc.

Marta M Meda

Geoffrey G Wilson

Ellen P Guthrie

Julie Forney Menin

The present invention is directed to a method for cloning and producing the HphI restriction endonuclease by 1) linking up the restriction gene to transcription elements that can be processed in E. coli and 2) expressing DNA modification enzymes in the host that protect against HphI digestion.

Isolated DNA encoding the hphi restriction endonuclease and related

The present invention is directed to a method for cloning and producing the SacII restriction endonuclease by 1) introducing the restriction endonuclease gene from Streptomyces achromogenes into a host whereby the restriction gene is expressed; 2) fermenting the host which contains the plasmid encoding and expressing the SacII restriction endonuclease activity, and 3) purifying the SacII restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the SacII restriction endonuclease activity.

Method for producing and cloning SacII restriction endonuclease and

The present invention provides a novel approach to the production of restriction enzymes. More specifically, there is provided a novel method for cloning these enzymes, which comprises preparing DNA libraries from the DNA of an organism that synthesizes the restriction enzyme of interest, creating a suitable host containing a heterospecific methyltransferase gene able to protectively modify DNA from digestion by the restriction enzyme of interest, introducing the DNA libraries into the protectively modified host, and screening recombinant organisms to identify those carrying the desired restriction enzyme gene. The application of this method to the FspI and HaeIII restriction genes of Fischerella species and Haemophilus aegyptius, respectively, is described in detail, together with the resulting clones that form the basis of a new and useful process for purifying the FspI and HaeIII restriction enzymes.

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Beverly, MA, United States of America

Marta M Meda