Zambrowicz B P, Friedrich G A, Buxton E C, Lilleberg S L, Person C, Sands A T
Lexicon Genetics, The Woodlands, Texas 77381, USA.
Nature. 1998 Apr 9;392(6676):608-11. doi: 10.1038/33423.
The dramatic increase in sequence information in the form of expressed sequence tags (ESTs) and genomic sequence has created a 'gene function gap' with the identification of new genes far outpacing the rate at which their function can be identified. The ability to create mutations in embryonic stem (ES) cells on a large scale by tagged random mutagenesis provides a powerful approach for determining gene function in a mammalian system; this approach is well established in lower organisms. Here we describe a high-throughput mutagenesis method based on gene trapping that allows the automated identification of sequence tags from the mutated genes. This method traps and mutates genes regardless of their expression status in ES cells. To facilitate the study of gene function on a large scale, we are using these techniques to create a library of ES cells called Omnibank, from which sequence-tagged mutations in 2,000 genes are described.
以表达序列标签（ESTs）和基因组序列形式存在的序列信息急剧增加，导致了“基因功能缺口”，新基因的识别速度远远超过其功能的识别速度。通过标记随机诱变在胚胎干细胞（ES细胞）中大规模产生突变的能力，为确定哺乳动物系统中的基因功能提供了一种强大的方法；这种方法在低等生物中已得到充分确立。在这里，我们描述了一种基于基因捕获的高通量诱变方法，该方法能够自动识别突变基因的序列标签。此方法可捕获并使基因发生突变，而不论其在ES细胞中的表达状态如何。为了便于大规模研究基因功能，我们正在使用这些技术创建一个名为全库（Omnibank）的ES细胞文库，从中描述了2000个基因的序列标签突变。
