Justice M J, Zheng B, Woychik R P, Bradley A
Life Sciences Division, Oak Ridge National Laboratory, Tennessee 37830, USA.
Methods. 1997 Dec;13(4):423-36. doi: 10.1006/meth.1997.0548.
The Human Genome Project has generated nucleotide sequences from an estimated 80,000 to 100,000 genes, only a small fraction of which have a known role. Nucleotide sequence information alone is insufficient to predict gene function. One of the most powerful ways of revealing gene function, as demonstrated in bacteria, worms, yeast, and flies, is to generate mutations and characterize them at both the phenotypic and the molecular levels. Given the physiological and anatomical parallels between mouse and human, genotype-phenotype relationships established in mice can be extrapolated to human syndromes. A new method is described for functional genetic analyses in the mouse that uses loxP/Cre engineering to generate coat color-tagged large deletions. The haploid regions can then be dissected by mutagenesis with N-ethyl-N-nitrosourea in phenotype-driven screens to obtain functional information on genes in any desired region of the mouse genome.
人类基因组计划已生成了约8万至10万个基因的核苷酸序列，其中只有一小部分的功能已知。仅核苷酸序列信息不足以预测基因功能。正如在细菌、线虫、酵母和果蝇中所证明的那样，揭示基因功能的最有效方法之一是产生突变并在表型和分子水平上对其进行表征。鉴于小鼠和人类在生理和解剖学上的相似性，在小鼠中建立的基因型-表型关系可以外推到人类综合征。本文描述了一种用于小鼠功能基因分析的新方法，该方法利用loxP/Cre工程产生带有毛色标记的大片段缺失。然后，可以通过在表型驱动的筛选中用N-乙基-N-亚硝基脲进行诱变来剖析单倍体区域，以获得小鼠基因组中任何所需区域基因的功能信息。
