Restriction endonuclease EcoO109 from Escherichia coli H709c with heptanucleotide recognition site 5'-PuG/GNCCPy |
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Authors: | K Mise K Nakajima |
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Affiliation: | 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 |
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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. |
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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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