Identification and characterisation of PmaCI an endonuclease of novel specificity from Pseudomonas maltophila.

Abstract:

: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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Walker JN,Dean PD,Saunders JR

doi

10.1093/nar/14.3.1293

subject

Has Abstract

pub_date

1986-02-11 00:00:00

pages

1293-301

issue

3

eissn

0305-1048

issn

1362-4962

journal_volume

14

pub_type

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