外显子捕获：一种用于在克隆的哺乳动物基因组DNA中鉴定候选转录序列的遗传筛选方法。
Exon trapping: a genetic screen to identify candidate transcribed sequences in cloned mammalian genomic DNA.
作者信息
Duyk G M, Kim S W, Myers R M, Cox D R
机构信息
Department of Pediatrics, University of California, San Francisco 94143-0554.
出版信息
Proc Natl Acad Sci U S A. 1990 Nov;87(22):8995-9. doi: 10.1073/pnas.87.22.8995.
Abstract
Identification and recovery of transcribed sequences from cloned mammalian genomic DNA remains an important problem in isolating genes on the basis of their chromosomal location. We have developed a strategy that facilitates the recovery of exons from random pieces of cloned genomic DNA. The basis of this "exon trapping" strategy is that, during a retroviral life cycle, genomic sequences of nonviral origin are correctly spliced and may be recovered as a cDNA copy of the introduced segment. By using this genetic assay for cis-acting sequences required for RNA splicing, we have screened approximately 20 kilobase pairs of cloned genomic DNA and have recovered all four predicted exons.
摘要
从克隆的哺乳动物基因组DNA中鉴定和回收转录序列，在基于基因的染色体定位来分离基因方面，仍然是一个重要问题。我们已经开发出一种策略，可促进从克隆的基因组DNA随机片段中回收外显子。这种“外显子捕获”策略的基础是，在逆转录病毒生命周期中，非病毒来源的基因组序列能够正确剪接，并可以作为导入片段的cDNA拷贝回收。通过使用这种针对RNA剪接所需顺式作用序列的遗传检测方法，我们已经筛选了约20千碱基对的克隆基因组DNA，并回收了所有四个预测的外显子。
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