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Restriction endonuclease EcoO109 from Escherichia coli H709c with heptanucleotide recognition site 5'-PuG/GNCCPy
Authors:K Mise  K Nakajima
Institution:1. Institute of Public Health, Shirokanedai 4-6-1, Minato-ku, Tokyo 108, Japan Tel. (03) 441-7111, ext. 264;2. Institute of Medical Sciences, Tokyo University, Shirokanedai 4-6-1, Minato-ku, Tokyo 108 Japan Tel. (03) 443-8111, ext. 343
Abstract:A new restriction endonuclease, EcoO109, has been isolated from Escherichia coli H709c by polyethyleneimine (PEI) precipitation, DEAE-cellulose chromatography and heparin agarose chromatography. The yield was high, more than 3000 units/g of wet cells. The EcoO109 endonuclease recognizes and cleaves a nucleotide sequence of (formula: see text), in the presence of 10 mM Mg2+. The enzyme will be useful for structural analysis and molecular cloning of DNA because of the stability, high yield and easy handling of the producer strain.
Keywords:Recombinant DNA  enzyme isolation and purification  DNA sequencing analysis  pBR322 vector  SV40 DNA  plasmids  Ad-2  adenovirus 2  bp  base pairs  DTT  1  4-dithiothreitol  host specificity for DNA  kb  1000 bp  nt  nucleotide(s)  PAB  Bacto-Pennassay broth (Difco)  PEI  polyethyleneimine  Pu  purine  Py  pyrimidine  RF  replicative form of DNA
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