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Nucleic Acids Research, 1975, Vol. 2, No. 10 1851-1866
© 1975

Articles

Sequences spanning the EcoRI substrate site

David E. Garfin, Herbert W. Boyer and Howard M. Goodman*

Departments of Microbiology and Biochemistry, University of California Medical Center San Francisco, CA 94143, USA *Department of Biophysics, University of California Medical Center San Francisco, CA 94143, USA

Received August 19, 1975. Substrate recognition by the EcoRI restriction endonuclease was investigated by analysis of the nucleotide sequences at the sites of enzymatic cleavage in various DNA molecules. 5'-end labeling and homochromatographic fingerprinting led to the determination of a 17-base-pair sequence spanning the EcoRI site of simian virus 40 DNA and a 15-base-pair sequence overlapping the EcoRI site of Col El plasmid DNA. Three other DNAs were similarly tested, although extended sequences were not determined in these cases. The EcoRI site was shown to be the symmetric, double-stranded equivalent of -N-G-A-A-T-T-C-N-.


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