Chumakov I M, Le Gall I, Billault A, Ougen P, Soularue P, Guillou S, Rigault P, Bui H, De Tand M F, Barillot E
Centre d'Etude du Polymorphisme Humain, Paris, France.
Nat Genet. 1992 Jun;1(3):222-5. doi: 10.1038/ng0692-222.
A new approach for the isolation of chromosome-specific subsets from a human genomic yeast artificial chromosome (YAC) library is described. It is based on the hybridization with an Alu polymerase chain reaction (PCR) probe. We screened a 1.5 genome equivalent YAC library of megabase insert size with Alu PCR products amplified from hybrid cell lines containing human chromosome 21, and identified a subset of 63 clones representative of this chromosome. The majority of clones were assigned to chromosome 21 by the presence of specific STSs and in situ hybridization. Twenty-nine of 36 STSs that we tested were detected in the subset, and a contig spanning 20 centimorgans in the genetic map and containing 8 STSs in 4 YACs was identified. The proposed approach can greatly speed efforts to construct physical maps of the human genome.
描述了一种从人类基因组酵母人工染色体（YAC）文库中分离染色体特异性亚群的新方法。该方法基于与Alu聚合酶链反应（PCR）探针的杂交。我们用从含人类21号染色体的杂交细胞系中扩增出的Alu PCR产物，筛选了一个插入片段大小为兆碱基的1.5倍基因组当量的YAC文库，并鉴定出代表该染色体的63个克隆的亚群。通过特定STS的存在和原位杂交，将大多数克隆定位到21号染色体。我们测试的36个STS中有29个在该亚群中被检测到，并且鉴定出一个在遗传图谱上跨越20厘摩、包含4个YAC中的8个STS的重叠群。所提出的方法可大大加快构建人类基因组物理图谱的工作。
