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Publication : A long-range regulatory element of Hoxc8 identified by using the pClasper vector.

First Author  Bradshaw MS Year  1996
Journal  Proc Natl Acad Sci U S A Volume  93
Issue  6 Pages  2426-30
PubMed ID  8637890 Mgi Jnum  J:32122
Mgi Id  MGI:79627 Doi  10.1073/pnas.93.6.2426
Citation  Bradshaw MS, et al. (1996) A long-range regulatory element of Hoxc8 identified by using the pClasper vector. Proc Natl Acad Sci U S A 93(6):2426-30
abstractText  Hox genes are located in highly conserved clusters. The significance of this organization is unclear, but one possibility is that regulatory regions for individual genes are dispersed throughout the cluster and shared with other Hox genes. This hypothesis is supported by studies on several Hox genes in which even large genomic regions immediately surrounding the gene fail to direct the complete expression pattern in transgenic mice. In particular, previous studies have identified proximal regulatory regions that are primarily responsible for early phases of mouse Hoxc8 expression. To locate additional regulatory regions governing expression during the later periods of development, a yeast homologous recombination-based strategy utilizing the pClasper vector was employed. Using homologous recombination into pClasper, we cloned a 27-kb region around the Hoxc8 gene from a yeast artificial chromosome. A reporter gene was introduced into the coding region of the isolated gene by homologous recombination in yeast. This large fragment recapitulates critical aspects of Hoxc8 expression in transgenic mice. We show that the regulatory elements that maintain the anterior boundaries of expression in the neural tube and paraxial mesoderm are located between 11 and 19 kb downstream of the gene.
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