Stark G R, Gudkov A V
Lerner Research Institute, The Cleveland Clinic Foundation, OH 44195, USA,
Hum Mol Genet. 1999;8(10):1925-38. doi: 10.1093/hmg/8.10.1925.
Definitive proof of function in biological systems requires genetic analysis. Only when the loss of a particular protein corresponds to the loss of a specific function can one be sure that the protein truly affects the function. Changing the pattern of gene expression through random mutagenesis or by introducing expression libraries, followed by selection of mutant or variant cells and identification of a missing or overexpressed protein, has the power to reveal or confirm the roles of specific components of signaling pathways and to provide mutant cell lines and cDNA reagents to be used in defining detailed mechanisms through structure-function analyses. These examples of forward genetics contrast with reverse genetic approaches, where the function of a known gene product is explored by knockout or replacement. Here we review a broad range of techniques that have been used to alter gene expression randomly in mammalian cells, with examples of specific discoveries that have resulted from these applications of forward genetics.
在生物系统中，功能的确切证据需要进行遗传分析。只有当特定蛋白质的缺失与特定功能的丧失相对应时，才能确定该蛋白质确实影响该功能。通过随机诱变或引入表达文库来改变基因表达模式，随后选择突变或变异细胞并鉴定缺失或过表达的蛋白质，有能力揭示或确认信号通路特定成分的作用，并提供突变细胞系和cDNA试剂，用于通过结构-功能分析来确定详细机制。这些正向遗传学的例子与反向遗传学方法形成对比，在反向遗传学方法中，通过基因敲除或替换来探索已知基因产物的功能。在这里，我们回顾了一系列已用于在哺乳动物细胞中随机改变基因表达的技术，并举例说明了这些正向遗传学应用所带来的具体发现。
