Artículos de revistas
Cracking the genome's second code: Enhancer detection by combined phylogenetic footprinting and transgenic fish and frog embryos
Fecha
2006Registro en:
Methods, Volumen 39, Issue 3, 2018, Pages 212-219
10462023
10959130
10.1016/j.ymeth.2005.12.005
Autor
Allende Connelly, Miguel
Manzanares, Miguel
Tena, Juan J.
Feijóo, Carmen G.
Gómez Skarmeta, José Luis
Institución
Resumen
Genes involved in vertebrate development are unusually enriched for highly conserved non-coding sequence elements. These regions are readily detected in silico, by genome-wide sequence comparisons between different vertebrates, from mammals to fish (phylogenetic footprinting). It follows that sequence conservation must be the result of positive selection for an essential physiological role. An obvious possibility is that these conserved sequences possess regulatory or structural functions important for gene expression and, thus, an in vivo assay becomes necessary. We have developed a rapid testing system using zebrafish and Xenopus laevis embryos that allows us to assign transcriptional regulatory functions to conserved non-coding sequence elements. The sequences are cloned into a vector containing a minimal promoter and the GFP reporter, and are assayed for their putative cis-regulatory activity in zebrafish or Xenopus transgenic experiments. Vectors used include plasmid DNA and the T