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Publication : Rapid identification of homeodomain binding sites in the Wnt-5a gene using an immunoprecipitation strategy.

First Author  Iler N Year  1996
Journal  Biochem Biophys Res Commun Volume  227
Issue  1 Pages  257-65
PubMed ID  8858134 Mgi Jnum  J:35778
Mgi Id  MGI:83223 Doi  10.1006/bbrc.1996.1498
Citation  Iler N, et al. (1996) Rapid identification of homeodomain binding sites in the Wnt-5a gene using an immunoprecipitation strategy. Biochem Biophys Res Commun 227(1):257-65
abstractText  Here we describe an immunoprecipitation approach for identifying homeodomain binding sites within uncharacterized genomic sequences of a putative downstream target gene, Wnt-5a. Immunoprecipitation of Wnt-5a genomic fragments was performed using a purified Msx1 homeodomain polypeptide (Msx1) and its corresponding antisera (alpha-Msx1). This resulted in isolation of three fragments containing multiple DNA binding sites for Msx1, as confirmed by DNA binding studies. The three fragments were contiguous within a 3.4 kb intronic sequence of Wnt-5a. Moreover, at least one of the Msx1 sites has been conserved throughout evolution, suggesting that these sites may comprise or contribute to a regulatory element for Wnt-5a. We propose that the immunoprecipitation strategy permits a rapid, initial approach for identifying functionally-relevant homeodomain binding sites within target genes whose regulatory sequences have not yet been previously elucidated.
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