Renault B, Hovnanian A, Bryce S, Chang J J, Lau S, Sakuntabhai A, Monk S, Carter S, Ross C J, Pang J, Twells R, Chamberlain S, Monaco A P, Strachan T, Kucherlapati R
Department of Molecular Genetics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
Genomics. 1997 Oct 15;45(2):271-8. doi: 10.1006/geno.1997.4888.
We developed a sequence-ready map of a part of human chromosome 12q24.1. We utilized a number of sequence-tagged site (STS) markers from 12q24.1 to screen large insert bacterial chromosome libraries and a chromosome 12-specific cosmid library. The clones were assembled into contiguous sets (contigs) by STS-content analysis. Contigs were extended by obtaining end sequences of bacterial clones, generation of additional STSs, rescreening the libraries, and screening the additional clones for the presence of STSs. The resulting contig covers nearly 2 Mb of DNA and provides an average marker resolution of 16 kb. Based on the STS content, we developed fingerprints of a subset of clones. The STS content and fingerprint data allowed us to define a minimal tiling path of clones. These clones are being used to sequence this part of chromosome 12. This contig contains the Ataxin 2 gene, and it covers the interval harboring the gene responsible for Darier disease.
我们绘制了人类12号染色体q24.1区域的序列就绪图谱。我们利用了来自12q24.1的多个序列标签位点(STS)标记来筛选大插入片段细菌染色体文库和12号染色体特异性黏粒文库。通过STS含量分析将克隆组装成连续片段(重叠群)。通过获取细菌克隆的末端序列、生成额外的STS、重新筛选文库以及筛选额外克隆中是否存在STS来扩展重叠群。最终得到的重叠群覆盖了近2 Mb的DNA,平均标记分辨率为16 kb。基于STS含量,我们绘制了一部分克隆的指纹图谱。STS含量和指纹数据使我们能够确定克隆的最小拼接路径。这些克隆正用于对12号染色体的这一区域进行测序。这个重叠群包含ataxin 2基因,并且覆盖了携带 Darier病致病基因的区间。