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Nucleic Acids Research, 1986, Vol. 14, No. 3 1293-1301
© 1986

Articles

Identification and characterisation of PmaCl an endonuclease of novel specificity from Pseudomonas maltophila

Jeremy N.B. Walker, Peter D.G. Dean+ and Jon R. Saunders

+ 34 Millington Road, Cambridge CB3 9PH, UK Department of Microbiology, Liverpool University P.O. Box 147, Liverpool L69 3BX, UK

Received October 31, 1985. Revised December 18, 1985. Accepted December 18, 1985.

We report the use of MonoQ FPLC (Fast Protein Liquid Chromatography) for the rapid purification of a novel Type II restriction endonuclease PmaCI, from Pseudomonas maltophila, which recognises the sequence 5'-CAC{downarrow}GTG-3'. The resulting enzyme is free of other nucleases to a level suitable for its characterisation by multiple-substrate digestion and DNA sequencing techniques. This method appears to be widely applicable and we have used it for the isolation of restriction endonucleases of comparable purity from a range of other organisms. Also described is a rapid method for screening a library of small inserted regions in recombinant M13 molecules for the presence and subsequent screening of restriction sites of interest.


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