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Nucleic Acids Research, 1985, Vol. 13, No. 24 8969-8981
© 1985

Articles

Interaction of EcoRII restriction and modification enzymes with synthetic DNA fragments. V. Study of single-strand cleavages

A.A. Yolov, E.S. Gromova, E.A. Kubareva, V.K. Potapov and Z.A. Shabarova

Department of Chemistry, Moscow State University Moscow 119899, USSR

Received August 6, 1985. Revised November 26, 1985. Accepted November 26, 1985.

Concatemer DNA duplexes which contain at the EcoRII restriction endonuclease cleavage sites Formula phospho-diester, phosphoamide or pyrophosphate internucleotide bonds have been synthesized. It has been shown that this enzyme did not cleave the substrate at phosphoamide bond. EcoRII endonuclease catalyzes single-strand cleavages both in dA- and dT-con-taining strands of the recognition site if the cleavage of the other strand has been blocked by modification of scissile bond or if the other strand has been cleaved. This enzyme interacts with both strands of the DNA recognition site, each of them being cleaved independently on the cleavage of another one. Nucleotide sequences flanking the EcoRII site on both sides are necessary for effective cleavage of the substrate.


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