Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1987-09-11
1989-02-28
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435 68, 435 70, 4351723, 435320, 435 6, 4353171, 935 1, 935 6, 935 10, 536 27, C12P 1934, C12P 2100, C07H 1512, C12Q 160
Patent
active
048085251
ABSTRACT:
The present invention provides a method for site-specific cleavage of double stranded DNA at sequences not less than eight base pairs long, comprising methylating he DNA with a sequence-specific methylase capable of recognizing and methylating a first sequence of the DNA, thereby generating a second sequence of the DNA capable of being recognized by a site-specific endonuclease, the first and second sequences having an overlapping part thereof; the length of the combined methylase and endonuclease recognition sites being not less than eight base pairs long, and cleaving the methylated DNA by treatment with the site-specific endonuclease. This method is useful for increasing the selectivity of cutting DNA stands using restriction endonucleases, thereby permitting the isolation of large DNA fragments and the generation of unique sites in DNA fragments, e.g., cloning vectors. Also provided is a DNA vector having a terminal sequence not less than eight base pairs long which can be recognized and cut by a site-specific endonuclease.
REFERENCES:
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"A DNA Methylase from Thermust Thermophilus HB8" J. Biochem 88, 737-747 (1980), Showbu Sato et al.
"Site-Specific cleavage of DNA at 8- and 10-Base-Pair Sequences" Proc. Nat'l. Acad. Sci., U.S.A. vol. 81, pp. 983-987, Feb. 1984 Biochemistry.
Nucleic Acids Research, vol. 9, No. 24 (1981), "Purification and Characterization of Two New Modification Methylases; . . . ".
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Kessler Louis G.
McClelland Michael
Seidman S.
University of Georgia Research Center
Wiseman Thomas G.
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