Moritz Orson L, Peck Allison, Tam Beatrice M
Department of Neuroscience, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-3401, USA
Gene. 2002 Oct 2;298(2):173-82. doi: 10.1016/s0378-1119(02)00923-x.
We have cloned the promoter regions of two Xenopus laevis genes, Prph2 (also called RDS) and red cone opsin (RCO) using a polymerase chain reaction-based gene-walking method. The proteins coded by these genes are expressed exclusively in retinal photoreceptors. Although these promoter sequences are evolutionarily distant from previously described homologues, potentially informative similarities were noted that suggest conserved binding sites of the transcription factors Crx and Rx. The promoters were tested for function in transgenic X. laevis. RCO-driven expression was restricted to cones and pinealocytes, while the Prph2 promoter drove expression of a reporter green fluorescent protein transgene in both rod and cone photoreceptors, as well as low levels of expression in muscle tissue. This is the first description of transgene expression driven by a Prph2 promoter homologue from any species. In combination with the previously reported X. laevis opsin and arrestin promoters, these sequences will facilitate the development and analysis of X. laevis models of inherited retinal degeneration.
我们使用基于聚合酶链反应的基因步移方法克隆了非洲爪蟾两个基因Prph2(也称为RDS)和红锥视蛋白(RCO)的启动子区域。这些基因编码的蛋白质仅在视网膜光感受器中表达。尽管这些启动子序列在进化上与先前描述的同源物相距甚远,但仍发现了潜在的有益相似性,提示转录因子Crx和Rx的保守结合位点。对这些启动子在转基因非洲爪蟾中的功能进行了测试。RCO驱动的表达仅限于视锥细胞和松果体细胞,而Prph2启动子则驱动报告基因绿色荧光蛋白转基因在视杆和视锥光感受器中表达,以及在肌肉组织中的低水平表达。这是首次描述来自任何物种的Prph2启动子同源物驱动的转基因表达。与先前报道的非洲爪蟾视蛋白和抑制蛋白启动子相结合,这些序列将有助于非洲爪蟾遗传性视网膜变性模型的开发和分析。