Loeber Jana, Pan Fong Cheng, Pieler Tomas
Department of Developmental Biochemistry, University of Goettingen, Goettingen, Germany.
Methods Mol Biol. 2009;561:65-72. doi: 10.1007/978-1-60327-019-9_4.
The possibility of generating transgenic animals is of obvious advantage for the analysis of gene function in development and disease. One of the established vertebrate model systems in developmental biology is the amphibian Xenopus laevis. Different techniques have been successfully applied to create Xenopus transgenics; in this chapter, the so-called meganuclease method is described. This technique is not only technically simple, but also comparably efficient and applicable to both Xenopus laevis and Xenopus tropicalis. The commercially available endonuclease I-SceI (meganuclease) mediates the integration of foreign DNA into the frog genome after coinjection into fertilized eggs. Tissue-specific gene expression, as well as germline transmission, has been observed.
生成转基因动物的可能性对于分析发育和疾病中的基因功能具有明显优势。发育生物学中已确立的脊椎动物模型系统之一是非洲爪蟾（Xenopus laevis）。不同的技术已成功应用于创建非洲爪蟾转基因动物；在本章中，将描述所谓的巨核酸酶方法。该技术不仅在技术上简单，而且效率相对较高，适用于非洲爪蟾和热带爪蟾。市售的内切核酸酶I-SceI（巨核酸酶）在与受精卵共注射后介导外源DNA整合到青蛙基因组中。已观察到组织特异性基因表达以及种系传递。
