Hamaguchi M, O'Connor E A, Chen T, Parnell L, McCombie R W, Wigler M H
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3764-9. doi: 10.1073/pnas.95.7.3764.
The isolation of genes from a given genomic region can be a rate-limiting step in the discovery of disease genes. We describe an approach to the isolation of cDNAs that have sequences in common with large genomic clones such as bacterial artificial chromosomes. We applied this method to loci both amplified and deleted in cancer, illustrating its usage in the identification of both oncogenes and tumor suppressor genes, respectively. The method, called rapid isolation of cDNAs by hybridization (RICH), depends on solution hybridization, enzymatic modification, and amplification/selection of sequences present in both cDNA populations and the genomic clones. The method should facilitate the development of transcription maps for large genomic clones, possibly even yeast artificial chromosomes.
从特定基因组区域分离基因可能是疾病基因发现过程中的一个限速步骤。我们描述了一种分离与大型基因组克隆（如细菌人工染色体）具有共同序列的cDNA的方法。我们将此方法应用于癌症中扩增和缺失的基因座，分别说明了其在鉴定癌基因和肿瘤抑制基因中的用途。该方法称为通过杂交快速分离cDNA（RICH），它依赖于溶液杂交、酶促修饰以及对cDNA群体和基因组克隆中都存在的序列进行扩增/选择。该方法应有助于大型基因组克隆甚至酵母人工染色体转录图谱的绘制。
