Abstract：
Transgenic approaches using I-SceI are powerful genome modification methods for creating heritable modifications in eukaryotic genomes. Such modifications are ideal for studying putative promoters and their temporal and spatial expression patterns in real time, in vivo. Central to this process is the initial engineering of a plasmid construct containing multiple DNA modules in a specific order prior to the integration into the target genome. One popular way of doing this is based upon the pGateway system, the modular form of which described in this chapter is known as pTransgenesis. We will initially describe the protocol of obtaining the plasmid construct containing the required sequence modules, and then the process of integrating the construct into the genome of a Xenopus embryo via co-injection with I-SceI and subsequent screening for transgenics.
摘要：
使用I-SceI的转基因方法是用于在真核基因组中产生可遗传修饰的强大基因组修饰方法。这种修饰对于实时研究推定的启动子及其时间和空间表达模式是理想的，在体内。该过程的核心是在整合到靶基因组之前以特定顺序对包含多个DNA模块的质粒构建体进行初始工程改造。一种流行的方法是基于pGateway系统，本章中描述的模块化形式称为pTransgenesis。我们将首先描述获得包含所需序列模块的质粒构建体的方案，然后通过与I-SceI共注射并随后筛选转基因，将构建体整合到非洲爪的胚胎基因组中。
