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Nucleic Acids Research, 1984, Vol. 12, No. 14 5567-5581
© 1984

SURVEYS AND SUMMARIES

A second type II restriction endonuclease from Thermus aquaticus with an unusual sequence specificity

David Barker*, Mark Hoff, Arnold Oliphant+ and Ray White

Howard Hughes Medical Institute, University of Utah Medical Center Salt Lake City, UT 84132, USA

Received June 18, 1984. Accepted June 29, 1984.

A Type II restriction endonuclease activity free of TaqI was prepared from Therraus Aquatious YT. The fraction contains two endonucleolytic components with apparently different specificities, however the major activity is sufficiently dominant to allow partial digestion analysis of the position of recognition sites. A precise determination of the location of cleavage sites in pBR322 DNA and a computer-aided search for regions of homology in the vicinity of the cut sites indicate that this enzyme recognizes the nonpalindromic sequences GACCGA or CACCCA. Other related sequences are not cleaved, in particular, GACCCA and CACCGA, indicating that the enzyme requires the identity of nucleotides in the first and fifth positions, a type of specificity that has not been previously reported. The position of cleavage is located outside of the site and is represented as:


Formula

*Present address: Collaborative Research Inc., 128 Spring Street, Lexington, MA 02173, USA

+Present address: Department of Genetics, Harvard Medical School, Boston, MA, USA


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