a multicassette gateway vector set for high throughput and comparative analyses in ciona and vertebrate embryosmulticassette网关向量集高吞吐量和比较分析ciona和脊椎动物的胚胎.pdfVIP

A Multicassette Gateway Vector Set for High Throughput and Comparative Analyses in Ciona and Vertebrate Embryos ` 1 ¨ 1 1 2 3 ´ 1 3 2 Agnes Roure *, Ute Rothbacher , Franc¸ois Robin , Eva Kalmar , Giustina Ferone , Clement Lamy , Caterina Missero , Ferenc Mueller , Patrick Lemaire1* ´ ´ ´ ´ 1 Institut de Biologie du Developpement de Marseille Luminy, UMR 6216 CNRS/Universite de la Mediterranee, Marseille, France, 2 Institute of Toxicology and Genetics, Forschungszentrum Karlsruhe, Karlsruhe, Germany, 3 CEINGE Biotecnologie Avanzate SCarl (Center for Genetic Engineering), Napoli, Italy 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. Principal Findings. 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. Conclusion. This versatile vector set opens the way to the mid- to large-scale comparative analyses of protein function and cis-regulatory sequences across ch