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

M. Suzanne Bradshaw, Cooduvalli S. Shashikant, Heinz Georg Belting, Jacques A. Bollekens, Frank H. Ruddle

Research output: Contribution to journalArticlepeer-review

52 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2426-2430
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number6
DOIs
StatePublished - Mar 19 1996

All Science Journal Classification (ASJC) codes

  • General

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