Improved Methods for Cloning Restriction Endonucleases

Current methods for cloning genes encoding restriction endonucleases are not efficient. This cloning process is relatively inefficient for two technical reasons: the preferred method is an indirect selection, selection for methylase activity, and the strength of the selection is only about 1000-fold. The specific aim is to optimize various features of a novel method by cloning BamHl. The long term objectives are to develop 5 to 10-fold more efficient methods for cloning restriction endonucleases. Cloned enzymes are usually cheaper and purer; therefore, the enzymes are more desirable for the research, clinical and industrial user. The methodology employed is a technological innovation potentially having additional health-related applications such as altering the specifity of naturally occurring restriction enzymes. More efficient cloning methods should interest both those commercial producers that are and those that are not currently making major efforts to clone these enzymes.