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Nucleic Acids Research, 1983, Vol. 11, No. 3 837-851
© 1983

MOLECULAR BIOLOGY

The Isolation and Characterization of the Esherichia Coli DNA adenine methylase (dam) gene

Joan E. Brooks, Robert M. Blumenthal+ and Thomas R. Gingeras

P.O. Box 100, Cold Spring Harbor Laboratory, Cold Spring Harbor Ny 11725, USA

Received October 18, 1982. Revised December 6, 1982. Accepted December 6, 1982.

The E coli dam (DNA adenine methylase) enxyne is known to nethylate the sequence GATC. A general nethod for cloning sequence-speoific DNA methylase genet was used to isolate the dam gene on a 1.14 kb fragment, inserted in the plasmid vector pBK322. Subsequent restriction mapping and subcloning experiments established a set of approximate boundaries of the gene. The nucleotide sequence of the dju gene was determined, and analysis of that sequence revealed a unique open reading frame which corresponded in length to that necessary to oode for a protein the sixe of dam. Amino acid conposition derived froa this sequence corresponds closely to the amino acid composition of the purified dam protein. Enxynatic and DNA:DNA hybridixation methods were used to investigate the possible presence of dasi genes in a variety of prokaryotlc organisms.

+Current address: Department of Microbiology, Medical College of Ohia Toledo, OH 43699, USA


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