Method of cleaving DNA

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

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435199, C12P 1934, C12N 922

Patent

active

054181500

ABSTRACT:
A method of cleaving substrate DNA with a restriction enzyme, wherein the substrate DNA is resistant to cleavage by the restriction enzyme, is disclosed. The method comprises co-incubating the substrate DNA and the restriction enzyme with an activating DNA sequence. The activating sequence comprises an oligonucleotide comprised of the restriction enzyme recognition site and cleavage permissible flanking sequences joined directly to both the 5' and 3' ends of the recognition site. Exemplary restriction enzymes which may be used in practicing the present invention include Nae I, BspM I, Hpa II, Nar I, and Sac II.

REFERENCES:
T. R. Gingeras and J. E. Brooks, Cloned restriction/modification system fromm Pseudomonas aeruginosa Proc. Natl. Acad. Sci. USA 80, 402-406 (1983).
D. H. Kruger et al. EcoRII can be activated to cleave refractory DNA recognition sites Nucleic Acids Research, 16, 3997-4008 (1988).
M. Conrad and Michael D. Topal, DNA and spermidine provide a switch mechanism to regulate the activity of restriction enzyme Nae I Proc. Natl. Acad. Sci. USA 86, 9707-9711. (1989).
C. D. Pein et al., Oligonucleotide duplexes containing CC(A/T)GG stimulate cleavage of refractory DNA by restriction endonuclease EcoRII FEBS Letters 245, 141-144 (1989).

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