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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
16
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pubmed:dateCreated |
1986-9-25
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pubmed:abstractText |
We describe a method for affinity purification of sequence-specific DNA binding proteins that is fast and effective. Complementary chemically synthesized oligodeoxynucleotides that contain a recognition site for a sequence-specific DNA binding protein are annealed and ligated to give oligomers. This DNA is then covalently coupled to Sepharose CL-2B with cyanogen bromide to yield the affinity resin. A partially purified protein fraction is combined with competitor DNA and subsequently passed through the DNA-Sepharose resin. The desired sequence-specific DNA binding protein is purified because it preferentially binds to the recognition sites in the affinity resin rather than to the nonspecific competitor DNA in solution. For example, a protein fraction that is enriched for transcription factor Sp1 can be further purified 500- to 1000-fold by two sequential affinity chromatography steps to give Sp1 of an estimated 90% homogeneity with 30% yield. In addition, the use of tandem affinity columns containing different protein binding sites allows the simultaneous purification of multiple DNA binding proteins from the same extract. This method provides a means for the purification of rare sequence-specific DNA binding proteins, such as Sp1 and CAAT-binding transcription factor.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-1100376,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-212715,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-2992804,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-2996137,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-3003068,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-4041012,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-4891970,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6095100,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6187469,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6225070,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6244545,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6273805,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6313230,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6597749,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-6987648,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3461465-7001362
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
83
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
5889-93
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
1986
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pubmed:articleTitle |
Affinity purification of sequence-specific DNA binding proteins.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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