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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
5
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pubmed:dateCreated |
1990-5-31
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pubmed:abstractText |
Endonuclease protection or "footprinting" analysis is a powerful technique for identifying the nucleotides involved in a protein-DNA interaction. DNase I is the most often employed endonucleolytic agent; however, this endonuclease does not exhibit the true nonsequence-specific cleavage desired for this type of analysis. Methidiumpropyl-EDTA (MPE) is a synthetic DNA intercalator that cleaves DNA in the presence of ferrous ion and oxygen. Cleavage by MPE exhibits no sequence specificity, a characteristic that makes this reagent better suited for protection analysis. Here we report a generally applicable technique for MPE protection (or "footprinting") analysis of specific DNA-protein complexes from a crude nuclear extract. We have used this method to identify the nucleotides of the immunoglobulin (Ig) heavy chain promoter region that are involved in complex formation with a protein that binds the octameric sequence ATGCAAAT, and we compare our results to those obtained previously using DNase I.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyribonuclease I,
http://linkedlifedata.com/resource/pubmed/chemical/Edetic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Iron Chelating Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/methidiumpropyl-EDTA-iron(II)
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
0736-6205
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
7
|
pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
500-4
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:2517395-Base Sequence,
pubmed-meshheading:2517395-Biotechnology,
pubmed-meshheading:2517395-Cell Nucleus,
pubmed-meshheading:2517395-DNA,
pubmed-meshheading:2517395-Deoxyribonuclease I,
pubmed-meshheading:2517395-Edetic Acid,
pubmed-meshheading:2517395-Iron Chelating Agents,
pubmed-meshheading:2517395-Molecular Sequence Data,
pubmed-meshheading:2517395-Proteins
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pubmed:year |
1989
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pubmed:articleTitle |
Protection analysis (or "footprinting") of specific protein-DNA complexes in crude nuclear extracts using methidiumpropyl-EDTA-iron (II).
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pubmed:affiliation |
University of Texas, Southwestern Medical Center.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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