Gossler A, Joyner A L, Rossant J, Skarnes W C
Mount Sinai Hospital Research Institute, Division of Molecular and Developmental Biology, Toronto, Canada.
Science. 1989 Apr 28;244(4903):463-5. doi: 10.1126/science.2497519.
A strategy was devised for identifying regions of the mouse genome that are transcriptionally active in a temporally and spatially restricted manner during development. The approach is based on the introduction into embryonic stem cells of two types of lacZ reporter constructs that can be activated by flanking mouse genomic sequences. Embryonic stem cells containing the lacZ constructs were used to produce chimaeric mice. Developmental regulation of lacZ expression occurred at a high frequency. Molecular cloning of the flanking endogenous genes and introduction of these potential insertional mutations into the mouse germ line should provide an efficient means of identifying and mutating novel genes important for the control of mammalian development.
设计了一种策略,用于识别小鼠基因组中在发育过程中以时间和空间受限方式转录活跃的区域。该方法基于将两种类型的lacZ报告基因构建体引入胚胎干细胞,这些构建体可被侧翼小鼠基因组序列激活。含有lacZ构建体的胚胎干细胞被用于产生嵌合小鼠。lacZ表达的发育调控以高频率发生。对侧翼内源基因进行分子克隆并将这些潜在的插入突变引入小鼠种系,应该为鉴定和突变对哺乳动物发育控制重要的新基因提供一种有效的方法。