Sible Jill C, Wroble Brian N
Department of Biological Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
Methods Mol Biol. 2009;518:1-15. doi: 10.1007/978-1-59745-202-1_1.
The microinjection of mRNA that is transcribed and capped in vitro into fertilized eggs and embryos of Xenopus laevis provides a powerful means for discovering the function of proteins during early development. Proteins may be overexpressed for a gain-of-function effect or exogenous protein function may be compromised by the microinjection of mRNA encoding "dominant-negative" proteins. This methodology is particularly suited for the investigation of the regulation of the cell cycle, checkpoints, and apoptosis in early development.
将体外转录并加帽的mRNA显微注射到非洲爪蟾的受精卵和胚胎中,为发现蛋白质在早期发育过程中的功能提供了一种强大的手段。蛋白质可通过过表达产生功能获得性效应,或者通过显微注射编码“显性负性”蛋白质的mRNA来破坏外源性蛋白质的功能。这种方法特别适合于研究早期发育过程中的细胞周期调控、检查点和细胞凋亡。