Identification and characterisation ofPmaCl an endonuclease of novel specificity fromPseudomonas maltophila | Zendy

Jeremy N.B. Walker | Zendy; P.D.G. Dean | Zendy; J. R. Saunders | Zendy

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Identification and characterisation ofPmaCl an endonuclease of novel specificity fromPseudomonas maltophila

Author(s) -

Jeremy N.B. Walker,

P.D.G. Dean,

J. R. Saunders

Publication year - 1986

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/14.3.1293

Subject(s) - biology , restriction enzyme , recombinant dna , endonuclease , fast protein liquid chromatography , microbiology and biotechnology , dna , enzyme , restriction digest , restriction map , ecori , biochemistry , restriction fragment , genetics , base sequence , gene

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 decreases 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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