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Titolo:
Exploring the DNA-binding specificities of zinc fingers with DNA microarrays
Autore:
Bulyk, ML; Huang, XH; Choo, Y; Church, GM;
Indirizzi:
Harvard Univ, Sch Med, Grad Biophys Program, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 ad Biophys Program, Boston, MA 02115 USA Harvard Univ, Sch Med, Dept Genet, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 ch Med, Dept Genet, Boston, MA 02115 USA MRC, Mol Biol Lab, Cambridge CB2 2QH, England MRC Cambridge England CB2 2QH , Mol Biol Lab, Cambridge CB2 2QH, England Gendaq Ltd, London NW7 1AD, England Gendaq Ltd London England NW7 1ADGendaq Ltd, London NW7 1AD, England
Titolo Testata:
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
fascicolo: 13, volume: 98, anno: 2001,
pagine: 7158 - 7163
SICI:
0027-8424(20010619)98:13<7158:ETDSOZ>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
HOMEODOMAIN RECOGNITION HELIX; PHAGE DISPLAY; PROTEIN; SELECTION; AFFINITY; SEQUENCES; COMPLEX; SYSTEM; SITES; CODE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
28
Recensione:
Indirizzi per estratti:
Indirizzo: Church, GM Harvard Univ, Sch Med, Grad Biophys Program, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 Program, Boston, MA 02115 USA
Citazione:
M.L. Bulyk et al., "Exploring the DNA-binding specificities of zinc fingers with DNA microarrays", P NAS US, 98(13), 2001, pp. 7158-7163

Abstract

A key step in the regulation of networks that control gene expression is the sequence-specific binding of transcription factors to their DNA recognition sites. A more complete understanding of these DNA-protein interactions will permit a more comprehensive and quantitative mapping of the regulatorypathways within cells, as well as a deeper understanding of the potential functions of individual genes regulated by newly identified DNA-binding sites. Here we describe a DNA microarray-based method to characterize sequence-specific DNA recognition by zinc-finger proteins. A phage display library,prepared by randomizing critical amino acid residues in the second of three fingers of the mouse Zif268 domain, provided a rich source of zinc-fingerproteins with variant DNA-binding specificities. Microarrays containing all possible 3-bp binding sites for the variable zinc fingers permitted the quantitation of the binding site preferences of the entire library, pools ofzinc fingers corresponding to different rounds of selection from this library, as well as individual Zif268 variants that were isolated from the library by using specific DNA sequences. The results demonstrate the feasibility of using DNA microarrays for genome-wide identification of putative transcription factor-binding sites.

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Documento generato il 25/11/20 alle ore 15:33:44