In situ detection of a heat-shock regulatory element binding protein using a soluble short synthetic enhancer sequence

In this study, a double stranded DNA sequence corresponding to the human heat shock regulatory element was chemically synthesized. By in vitro retardation assays, the synthetic sequence was shown to bind specifically a protein in extracts from the human T cell line Jurkat. When the synthetic enhance...

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Bibliographic Details
Published in:Nucleic acids research 1989-01, Vol.17 (11), p.4077-4087
Main Authors: HAREL-BELLAN, A, BRINI, A. T, FERRIS, D. K, ROBIN, P, FARRAR, W. L
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Binding and carrier proteins
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Proteins
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Summary:In this study, a double stranded DNA sequence corresponding to the human heat shock regulatory element was chemically synthesized. By in vitro retardation assays, the synthetic sequence was shown to bind specifically a protein in extracts from the human T cell line Jurkat. When the synthetic enhancer was electroporated into Jurkat cells, not only the enhancer was shown to remain undegraded into the cells for up to 2 days, but also it was shown to bind intracellularly a protein. The binding was specific and was modulated upon heat shock. Futhermore, the binding protein was shown to be of the expected molecular weight by UV crosslinking.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/17.11.4077