Publication Date:
1997-01-31
Description:
A method is described for selecting DNA-binding proteins that recognize desired sequences. The protocol involves gradually extending a new zinc finger protein across the desired 9- or 10-base pair target site, adding and optimizing one finger at a time. This procedure was tested with a TATA box, a p53 binding site, and a nuclear receptor element, and proteins were obtained that bind with nanomolar dissociation constants and discriminate effectively (greater than 20,000-fold) against nonspecific DNA. This strategy may provide important information about protein-DNA recognition as well as powerful tools for biomedical research.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Greisman, H A -- Pabo, C O -- New York, N.Y. -- Science. 1997 Jan 31;275(5300):657-61.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Howard Hughes Medical Institute and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/9005850" target="_blank"〉PubMed〈/a〉
Keywords:
Amino Acid Sequence
;
Base Composition
;
Base Sequence
;
Binding Sites
;
DNA/*metabolism
;
DNA-Binding Proteins/chemistry/*metabolism
;
Genes, p53
;
Hydrogen Bonding
;
Models, Molecular
;
Molecular Sequence Data
;
Nucleic Acid Conformation
;
Peptide Library
;
Protein Conformation
;
*Protein Engineering
;
Protein Structure, Secondary
;
Receptors, Cytoplasmic and Nuclear/genetics
;
TATA Box
;
Transcription Factors/chemistry/metabolism
;
*Zinc Fingers
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
