Haeri Mohammad, Knox Barry E
Department of Neuroscience & Physiology, SUNY Upstate Medical University, Syracuse, NY, USA.
Methods Mol Biol. 2012;884:17-39. doi: 10.1007/978-1-61779-848-1_2.
Transgenesis, the process of incorporating an exogenous gene (transgene) into an organism's genome, is a widely used tool to develop models of human diseases and to study the function and/or regulation of genes. Generating transgenic Xenopus is rapid and involves simple in vitro manipulations, taking advantage of the large size of the amphibian egg and external embryonic development. Restriction enzyme-mediated integration (REMI) has a number of advantages for transgenesis compared to other methods used to produce transgenic Xenopus, including relative efficiency, higher transgene expression levels, fewer genetic chimera in founder transgenic animals, and near-complete germ-line transgene transmission. This chapter explains the REMI method for generating transgenic Xenopus laevis tadpoles, including improvements developed to enable studies in the mature retina.
转基因技术是将外源基因（转基因）整合到生物体基因组中的过程，是开发人类疾病模型以及研究基因功能和/或调控的广泛应用工具。利用两栖类卵细胞体积大及胚胎体外发育的特点，生成转基因非洲爪蟾快速且只需简单的体外操作。与用于产生转基因非洲爪蟾的其他方法相比，限制性内切酶介导整合（REMI）在转基因方面具有许多优势，包括相对效率、更高的转基因表达水平、转基因动物始祖中较少的遗传嵌合体以及近乎完全的种系转基因传递。本章阐述了用于生成转基因非洲爪蟾蝌蚪的REMI方法，包括为在成熟视网膜中开展研究而进行的改进。
