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Nucleic Acids Research, 1985, Vol. 13, No. 2 389-399
© 1985

Articles

The nucleotide sequence recognised by the Escherichia coli D type I restriction and modification enzyme

Valakunja Nagaraja, Martin Stieger, Christoph Nager, Sheikh M. Hadi and Thomas A. Bickle*

Microbiology Department, Biozentrum, University of Basel Klingelbergstrasse 70, CH-4056 Basel, Switzerland

*To whom correspondance should be addressed

Received October 24, 1984. Revised December 31, 1984. Accepted December 31, 1984.

A type I restriction endonuclease from a new isolate of Escherichia coli (E. coli E166) has been purified and characterised. The enzyme, EcoD, has a recognition sequence similar in overall structure to the previously determined type I enzyme sequences, an exception being that it is degenerate. The sequence is

5'-T-T-A-N-N-N-N-N-N-N-G-T-C-Y-3'

3'-A-A-T-N-N-N-N-N-N-N-C-A-G-R-5'

where Y is a pyrimidine, R is a purine and N can be any nucleotide. The enzyme methylates adenosyl residues in both strands of the DNA that are separated by ten base pairs, suggesting that the enzyme interacts with DNA along one face of the helix making contacts in two successive major grooves.


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