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Nucleic Acids Research, 1982, Vol. 10, No. 3 993-1007
© 1982

ENZYMOLOGY

Preferential site-dependent cleavage by restriction endonuclease PstI

Karen Armstrong and William R. Bauer

Department of Microbiology, School of Medicine, State University of New York at Stony Brook Stony Brook, NY 11794, USA

Received October 12, 1981. Revised December 21, 1981. Accepted December 21, 1981.

The four identical recognition sites for the restriction endonuclease PstI in purified plasmid pSMl DNA I are cleaved at markedly different rates. The order and relative frequencies of cleavage at these four PstI sites have been determined from the order of appearance of partial cleavage products and from an analysis of production of specific unit length linear molecules. The same pattern of preferential cleavage is also found when linear, nicked circular, or relaxed closed circular forms of the same plasmid DNA are used as substrates for PstI. Inspection of the nucleotide sequences immediately adjoining each of the PstI sites suggests that the presence of adjacent runs of G-C base pairs confers significant resistance to cleavage.


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