The Isolation and Characterization of theEsherichia ColiDNA adenine methylase(dam)gene | Zendy

Joan E. Brooks | Zendy; Robert Blumenthal | Zendy; T Gingeras | Zendy

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The Isolation and Characterization of theEsherichia ColiDNA adenine methylase(dam)gene

Author(s) -

Joan E. Brooks,

Robert Blumenthal,

T Gingeras

Publication year - 1983

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/11.3.837

Subject(s) - biology , spring (device) , isolation (microbiology) , dna , library science , art history , genetics , history , bioinformatics , physics , computer science , thermodynamics

The E. coli dam (DNA adenine methylase) enzyme is known to methylate the sequence GATC. A general method for cloning sequence-specific DNA methylase genes was used to isolate the dam gene on a 1.14 kb fragment, inserted in the plasmid vector pBR322. Subsequent restriction mapping and subcloning experiments established a set of approximate boundaries of the gene. The nucleotide sequence of the dam gene was determined, and analysis of that sequence revealed a unique open reading frame which corresponded in length to that necessary to code for a protein the size of dam. Amino acid composition derived from this sequence corresponds closely to the amino acid composition of the purified dam protein. Enzymatic and DNA:DNA hybridization methods were used to investigate the possible presence of dam genes in a variety of prokaryotic organisms.

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