Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
21
pubmed:dateCreated
1991-1-3
pubmed:abstractText
We describe a sensitive and rapid method for determination of the sequence specificity of DNA binding proteins. The method allows recovery of specific sites using the small amounts of protein present in crude cell extracts or produced by cell-free translation reactions. Extract proteins are incubated with a pool of random sequence oligonucleotides, complexes purified by immunoprecipitation, and bound DNA amplified by the Polymerase Chain Reaction (PCR). This DNA is then used in further rounds of binding, immunoprecipitation, and amplification, until specific binding is detectable. With the transcription factor SRF as a model system, we demonstrate that authentic high affinity binding sites are recovered, and show that epitope tagging can be used to allow recovery of sites when specific antibodies are unavailable. We also show that specific sites bound by the Fos protein, which binds DNA with high affinity only when complexed with other polypeptides, are easily recovered by this technique.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-1969796, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2105456, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2110922, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2133110, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2159142, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2192357, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2414012, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2415984, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2423876, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2455264, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2493627, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2494661, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2503252, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2504586, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2507923, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2510170, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2550138, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2571934, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2673922, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2674675, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2734098, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2797150, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2797151, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2904838, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2908196, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-2999980, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3046753, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3112583, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3119326, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3133553, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3141060, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3185713, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3203386, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3282223, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3323803, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3323821, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3524858, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3545499, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-3915782, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-4079979, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-6258090, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-6267291, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-6526011, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-6656641, http://linkedlifedata.com/resource/pubmed/commentcorrection/2243767-6828386
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0305-1048
pubmed:author
pubmed:issnType
Print
pubmed:day
11
pubmed:volume
18
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
6197-204
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1990
pubmed:articleTitle
A sensitive method for the determination of protein-DNA binding specificities.
pubmed:affiliation
Imperial Cancer Research Fund Laboratories, Lincoln's Inn Fields, London, UK.
pubmed:publicationType
Journal Article