Marykwas D L, Passmore S E
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11701-5. doi: 10.1073/pnas.92.25.11701.
An efficient method for mapping mutations is described in which hybrid genes, derived partly from mutant and partly from wild-type DNA, are obtained in vivo by homologous recombination of multiple fragments. The recombinants are formed in a strain in which their phenotypes are immediately apparent. This method was developed to identify changes that disrupt protein-protein interactions demonstrable by the two-hybrid system in yeast. However, it can be extended to any system where recombination is possible, provided an assay is available to distinguish between mutant and wild-type phenotypes.
本文描述了一种高效的突变定位方法，该方法通过多个片段的同源重组在体内获得部分源自突变体DNA、部分源自野生型DNA的杂交基因。重组体在其表型能立即显现的菌株中形成。开发此方法是为了识别那些破坏酵母双杂交系统中可证明的蛋白质-蛋白质相互作用的变化。然而，只要有检测方法可区分突变型和野生型表型，该方法就可扩展到任何可能发生重组的系统。
