Generating transgenic frog embryos by restriction enzyme mediated integration (REMI).
Generating transgenic frog embryos by restriction enzyme mediated integration (REMI).
复制标题
DOI：
10.1007/978-1-61779-992-1_11
复制
发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Amaya, Enrique
中科院分区：
文献类型：
--
作者：
Ishibashi, Shoko;Kroll, Kristen L;Amaya, Enrique
在这里，我们提出了一种在非洲爪哇和热带爪哇中产生转基因胚胎的方法。该方法包括三个步骤：(1)制备高速卵子提取物，促进精子核中的鱼精蛋白被核小体取代，并解聚精子核的染色质；(2)精子核的分离；(3)精子核、限制性内切酶和体外高速提取物的混合，然后核移植到未受精卵中产生转基因胚胎。这一过程产生了高频率和高效率的非镶嵌转基因胚胎。
Here we present a protocol for generating transgenic embryos in Xenopus laevis and Xenopus tropicalis. The method includes three steps: (1) The preparation of high-speed egg extracts, which facilitates the replacement of protamines in sperm nuclei with nucleosomes and decondenses the chromatin of sperm nuclei; (2) The isolation of sperm nuclei; and (3) The mixing of sperm nuclei, restriction enzyme, and high-speed extract in vitro, following by nuclear transplantation into unfertilized eggs to generate the transgenic embryos. This procedure generates non-mosaic transgenic embryos at high frequency and efficiency.