title: Manipulation of single-stranded DNA by using an artificial site-selective DNA cutter composed of cerium(IV)/EDTA and phosphonate-oligonucleotide conjugates.
abstract: An artificial site-selective DNA cutter to hydrolyze single-stranded DNA at a desired site was prepared from Ce(IV)/ethylenediamintetraacetic acid (EDTA) and two ethylenediamine-N,N,N',N'-tetrakis(methylenephosphonic acid)-oligonucleotide conjugates. By using this cutter, the sense strand of a blue fluorescent protein (BFP) gene was selectively cut at a predetermined site in the chromophore-coding region. The upstream fragment obtained by the site-selective scission was ligated with the downstream fragment of the closely related green fluorescent protein (GFP) gene so that the 5'- and 3'-end portions of the chromophore came from the BFP fragment and the GFP fragment, respectively. The recombinant gene was successfully expressed in E. coli and the chimeric chromophore emitted green fluorescence as expected.
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Aiba
authors.forename: Tuomas
authors.initials: T
authors.lastname: Lönnberg
authors.forename: Makoto
authors.initials: M
authors.lastname: Komiyama
mesh_terms: D002563:Cerium / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D004492:Edetic Acid / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D006868:Hydrolysis; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry*; D063065:Organophosphonates / Q000737:chemistry*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004277:DNA, Single-Stranded; D008164:Luminescent Proteins; D009841:Oligonucleotides; D063065:Organophosphonates; D011994:Recombinant Proteins; D002563:Cerium; D004492:Edetic Acid