Swan Kathryn A, Curtis Damian E, McKusick Kathleen B, Voinov Alexander V, Mapa Felipa A, Cancilla Michael R
Exelixis, Inc., South San Francisco, California 94083-0511, USA.
Genome Res. 2002 Jul;12(7):1100-5. doi: 10.1101/gr.208902.
Positional cloning of mutations in model genetic systems is a powerful method for the identification of targets of medical and agricultural importance. To facilitate the high-throughput mapping of mutations in Caenorhabditis elegans, we have identified a further 9602 putative new single nucleotide polymorphisms (SNPs) between two C. elegans strains, Bristol N2 and the Hawaiian mapping strain CB4856, by sequencing inserts from a CB4856 genomic DNA library and using an informatics pipeline to compare sequences with the canonical N2 genomic sequence. When combined with data from other laboratories, our marker set of 17,189 SNPs provides even coverage of the complete worm genome. To date, we have confirmed >1099 evenly spaced SNPs (one every 91 +/- 56 kb) across the six chromosomes and validated the utility of our SNP marker set and new fluorescence polarization-based genotyping methods for systematic and high-throughput identification of genes in C. elegans by cloning several proprietary genes. We illustrate our approach by recombination mapping and confirmation of the mutation in the cloned gene, dpy-18.
在模式遗传系统中对突变进行定位克隆是一种用于鉴定具有医学和农业重要性靶点的强大方法。为了便于对秀丽隐杆线虫中的突变进行高通量定位，我们通过对来自CB4856基因组DNA文库的插入片段进行测序，并使用信息学流程将序列与标准的N2基因组序列进行比较，在两种秀丽隐杆线虫菌株（布里斯托尔N2和夏威夷定位菌株CB4856）之间鉴定出另外9602个推定的新单核苷酸多态性（SNP）。当与其他实验室的数据相结合时，我们的17189个SNP标记集能够均匀覆盖完整的线虫基因组。迄今为止，我们已经在六条染色体上确认了超过1099个均匀分布的SNP（每91±56 kb一个），并通过克隆几个专利基因验证了我们的SNP标记集和基于新的荧光偏振的基因分型方法在秀丽隐杆线虫中系统且高通量鉴定基因的实用性。我们通过重组定位和对克隆基因dpy-18中的突变进行确认来说明我们的方法。
