Source:http://linkedlifedata.com/resource/pubmed/id/15538776
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
22
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pubmed:dateCreated |
2004-11-12
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pubmed:abstractText |
In eukaryotes, transcription is regulated by multiprotein complexes binding to specific regions of genomic DNA, called cis-regulatory elements. Comprehensive identification of these elements is an important goal of functional genomics. Hence, it is of practical interest to develop a high-throughput assay to identify cis-regulatory elements. Toward that goal, we demonstrate that a surface plasmon resonance-based assay can identify whether a specific region of DNA binds to proteins present in raw nuclear lysate. Specifically, we immobilized a 16-basepair double-stranded DNA region of the SQSTM1 promoter to the Texas Instruments Spreeta, a surface plasmon resonance sensor. As a control, in a separate experiment, we immobilized a similar piece of DNA that differed by only a single base pair. We observed a significant difference in surface plasmon resonance signal when these two probes were exposed to raw nuclear lysate from NIH/3T3 cells. Using a luciferase-reporter vector transfected into live NIH/3T3 cells, we measured a significant difference in transcriptional activity between the two pieces of DNA. We conclude that a surface plasmon resonance-based sensor is capable of identifying physiologically significant cis-regulatory elements.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0003-2700
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
76
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
6555-9
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pubmed:dateRevised |
2009-1-19
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pubmed:meshHeading |
pubmed-meshheading:15538776-Adaptor Proteins, Signal Transducing,
pubmed-meshheading:15538776-Adsorption,
pubmed-meshheading:15538776-Animals,
pubmed-meshheading:15538776-Mice,
pubmed-meshheading:15538776-NIH 3T3 Cells,
pubmed-meshheading:15538776-Promoter Regions, Genetic,
pubmed-meshheading:15538776-Proteins,
pubmed-meshheading:15538776-Regulatory Sequences, Nucleic Acid,
pubmed-meshheading:15538776-Surface Plasmon Resonance
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pubmed:year |
2004
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pubmed:articleTitle |
Surface plasmon resonance-based sensors to identify cis-regulatory elements.
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pubmed:affiliation |
Department of Biomedical Engineering, University of California-Irvine, Irvine, California 92697-2715, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, N.I.H., Extramural
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