Ooi S L, Shoemaker D D, Boeke J D
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, 617 Hunterian Building, 725 North Wolfe Street, Baltimore, MD 21205, USA.
Science. 2001 Dec 21;294(5551):2552-6. doi: 10.1126/science.1065672. Epub 2001 Nov 8.
We describe a microarray-based screen performed by imposing different genetic selections on thousands of yeast mutants in parallel, representing most genes in the yeast genome. The presence or absence of mutants was detected by oligonucleotide arrays that hybridize to 20-nucleotide "barcodes." We used this method to screen for components of the nonhomologous end-joining (NHEJ) pathway. Known components of the pathway were identified, as well as a gene not previously known to be involved in NHEJ, NEJ1. Nej1 protein interacts with the amino terminus of LIF1/XRCC4, a recently recognized "guardian of the genome" against cancer.
我们描述了一种基于微阵列的筛选方法，该方法通过对数千个酵母突变体同时施加不同的遗传选择来进行，这些突变体代表了酵母基因组中的大多数基因。通过与20个核苷酸的“条形码”杂交的寡核苷酸阵列来检测突变体的存在或缺失。我们使用这种方法筛选非同源末端连接（NHEJ）途径的组成成分。该途径的已知组成成分被鉴定出来，同时还发现了一个以前未知参与NHEJ的基因NEJ1。Nej1蛋白与LIF1/XRCC4的氨基末端相互作用，LIF1/XRCC4是最近被认可的一种抗癌“基因组守护者”。
