A gene trap approach in Xenopus
A gene trap approach in Xenopus
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DOI：
10.1016/s0960-9822(00)80025-1
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发表时间：
1999-10-21
期刊：
影响因子：
9.2
通讯作者：
Amaya, E
中科院分区：
文献类型：
--
作者：
Bronchain, OJ;Hartley, KO;Amaya, E
青蛙转基因技术最终有望通过随机插入质粒DNA到基因组中使诱变成为可能。本研究旨在评估基因诱捕与转基因相结合的方法是否适用于非洲爪蟾胚胎的插入诱变。首先，我们证实了转基因技术可以稳定地整合到基因组中，并且通过种系传播以预期的孟德尔方式发生。其次，以绿色荧光蛋白（GFP）为标记，构建了多种基因诱捕载体。利用这些载体，我们捕获了非洲爪蟾中以空间受限方式表达的几个基因，包括骨骺、嗅球和基板、眼睛、耳朵、大脑、肌肉、尾巴和肠道中的表达。最后，我们利用5′快速扩增cDNA末端聚合酶链反应（RACE PCR）克隆了其中一个捕获基因，这些结果表明，转基因技术与基因诱捕方法相结合，可能为爪蟾内源基因突变的产生提供了一种有效的方法。
The frog transgenesis technique ultimately promises to make mutagenesis possible through random insertion of plasmid DNA into the genome. This study was undertaken to evaluate whether a gene trap approach combined with transgenesis would be appropriate for performing insertional mutagenesis in Xenopus embryos. Firstly, we confirmed that the transgenic technique results in stable integration into the genome and that transmission through the germline occurs in the expected Mendelian fashion. Secondly, we developed several gene trap vectors, using the green fluorescent protein (GFP) as a marker. Using these vectors, we trapped several genes in Xenopus laevis that are expressed in a spatially restricted manner, including expression in the epiphysis, the olfactory bulb and placodes, the eyes, ear, brain, muscles, tail and intestine. finally, we cloned one of the trapped genes using 5' rapid amplification of cDNA ends polymerase chain reaction (RACE PCR), These results suggest that the transgenic technique combined with a gene trap approach might provide a powerful method for generating mutations in endogenous genes in Xenopus.