A multicassette Gateway vector set for high throughput and comparative analyses in ciona and vertebrate embryos.
BACKGROUND
The past few years have seen a vast increase in the amount of genomic data available for a growing number of taxa, including sets of full length cDNA clones and cis-regulatory sequences. Large scale cross-species comparisons of protein function and cis-regulatory sequences may help to understand the emergence of specific traits during evolution.


RESULTS
To facilitate such comparisons, we developed a Gateway compatible vector set, which can be used to systematically dissect cis-regulatory sequences, and overexpress wild type or tagged proteins in a variety of chordate systems. It was developed and first characterised in the embryos of the ascidian Ciona intestinalis, in which large scale analyses are easier to perform than in vertebrates, owing to the very efficient embryo electroporation protocol available in this organism. Its use was then extended to fish embryos and cultured mammalian cells.


CONCLUSIONS
This versatile vector set opens the way to the mid- to large-scale comparative analyses of protein function and cis-regulatory sequences across chordate evolution. A complete user manual is provided as supplemental material.
Roure
Agnès
A
Institut de Biologie du Développement de Marseille Luminy, UMR 6216 CNRS/Université de la Méditerranée, Marseille, France. roure@ibdm.univ-mrs.fr
Rothbächer
Ute
U
Robin
François
F
Kalmar
Eva
E
Ferone
Giustina
G
Lamy
Clément
C
Missero
Caterina
C
Mueller
Ferenc
F
Lemaire
Patrick
P
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002938:Ciona intestinalis / Q000196:embryology*; D017931:DNA Primers; D018076:DNA, Complementary; D016366:Open Reading Frames; D012045:Regulatory Sequences, Nucleic Acid; D014714:Vertebrates / Q000196:embryology*
D017931:DNA Primers; D018076:DNA, Complementary