Process for producing recombinant McrBC endonuclease and cleavag

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

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435199, 43525233, 4353201, 536 232, C12N 922, C12N 1555, C12N 1570

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active

054057604

ABSTRACT:
The present invention relates to a recombinant McrBC endonuclease obtainable from Escherichia coli, two components of which, McrB.sub.L and McrC, have been purified in active form. McrBC is active in the presence of GTP and at a low pH. The McrBC endonuclease is also substantially free of a third component, McrB.sub.S, which is believed to inhibit or otherwise interfere with the activity of the enzyme. McrBC has various desirable properties, including the ability to recognize a methylated DNA sequence and also its ability to cleave such a sequence in the presence of GTP. Also provided is a process for the production of recombinant McrBC endonuclease, a process for the determination of the modification state of DNA a process for the determination of an epigenetic alteration or defect (including "imprinting"), as well as a process for identifying and isolating additional enzymes that cleave modified DNA.

REFERENCES:
patent: 5082784 (1992-01-01), Chatterjee et al.
Sutherland, E. et al., (1992), J. Mol. Biol., 225, 327-348.
Zheng, L., et al., (1992), Gene, p. 112, 97-100.

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