Schön Christian, Köster Manfred, Knöchel Walter
Abteilung Biochemie, Universität Ulm, Albert-Einstein-Allee 11, D-89081 Ulm, Germany.
Biochem Biophys Res Commun. 2004 Dec 24;325(4):1360-6. doi: 10.1016/j.bbrc.2004.10.177.
We have isolated and sequenced the pseudo-allelic versions of FoxD5 genes in Xenopus laevis, xlFoxD5a and xlFoxD5b, and the xtFoxD5 gene of Xenopus tropicalis. These genes show a highly conserved structure, they are composed of only one exon, and they exhibit a high degree of sequence conservation within their flanking sequences. The X. tropicalis gene is expressed like its X. laevis orthologues in progenitor cells of the neural floor plate. Serial deletions of the 5'- and 3'-flanking region in combination with reporter gene assays demonstrate that a distal and a proximal upstream element as well as a downstream-located enhancer do mainly contribute to transcriptional activity. The downstream enhancer cooperates with the proximal upstream element and also contributes to the spatial expression. Transgenic animals express enhanced green fluorescent protein in a spatial pattern like xlFoxD5b, when the 5'- and 3'-flanking regions of the xlFoxD5b gene are used to direct transgene expression.
我们已经分离并测序了非洲爪蟾(Xenopus laevis)中FoxD5基因的拟等位基因版本xlFoxD5a和xlFoxD5b,以及热带爪蟾(Xenopus tropicalis)的xtFoxD5基因。这些基因显示出高度保守的结构,它们仅由一个外显子组成,并且在其侧翼序列内表现出高度的序列保守性。热带爪蟾基因在神经底板的祖细胞中与其非洲爪蟾直系同源基因一样表达。5'和3'侧翼区域的系列缺失与报告基因检测相结合表明,一个远端和一个近端上游元件以及一个位于下游的增强子主要对转录活性有贡献。下游增强子与近端上游元件协同作用,也有助于空间表达。当使用xlFoxD5b基因的5'和3'侧翼区域来指导转基因表达时,转基因动物以类似于xlFoxD5b的空间模式表达增强型绿色荧光蛋白。