Institut de Biologie du Développement de Marseille Luminy, UMR 6216 CNRS/Université de la Méditerranée, Marseille, France.
PLoS One. 2007 Sep 19;2(9):e916. doi: 10.1371/journal.pone.0000916.
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.
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.
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.
在过去的几年中，可供越来越多分类单元使用的基因组数据数量大幅增加，包括全长 cDNA 克隆和顺式调控序列集。对蛋白质功能和顺式调控序列进行大规模跨物种比较，可能有助于理解进化过程中特定特征的出现。
为了便于进行此类比较，我们开发了一套兼容 Gateway 的载体系统，可用于系统地剖析顺式调控序列，并在多种脊索动物系统中过表达野生型或标记蛋白。该系统最初在尾索动物海鞘的胚胎中进行了开发和特征描述，由于该生物中存在非常有效的胚胎电穿孔技术，因此在该系统中进行大规模分析比在脊椎动物中更容易。随后，该系统的应用范围扩展到鱼类胚胎和培养的哺乳动物细胞。
该多功能载体系统为跨脊索动物进化进行蛋白质功能和顺式调控序列的中大规模比较分析开辟了道路。我们提供了完整的用户手册作为补充材料。
