Hubert R S, Mitchell S, Chen X N, Ekmekji K, Gadomski C, Sun Z, Noya D, Kim U J, Chen C, Shizuya H, Simon M, de Jong P J, Korenberg J R
Abmanson Department of Pediatrics, CSMC Burns and Allen Research Institute, Los Angeles, California, USA.
Genomics. 1997 Apr 15;41(2):218-26. doi: 10.1006/geno.1997.4657.
Chromosome 21 is a model for the study of human chromosomal aneuploidy, and the construction of its physical and transcriptional maps is a necessary step in understanding the molecular basis of aneuploidy-dependent phenotypes. To identify the gene(s) responsible for Down syndrome congenital heart disease (DS-CHD), we constructed a physical map of the D21S55 to MX1 region. A bacterial artificial chromosome (BAC) library was screened using several YACs spanning the interval, and a P1-derived artificial chromosome (PAC) library was screened using radiolabeled STS PCR products and whole BACs in gap-filling initiatives. FISH confirmed the location of all BAC and PAC clones to 21q22.2-q22.3. Overlaps were established using clone-to-clone Southerns and 24 new STSs, generated from the direct sequencing of BAC and PAC ends, along with 35 preexisting STSs. Approximately 3.5 Mb of the 4- to 5-Mb D21S55 to MX1 interval is covered in 85 BACs and 24 PACs, representing fourfold coverage within the contigs. These BAC and PAC contigs are valuable reagents for isolating the genes for DS-CHD.
21号染色体是研究人类染色体非整倍性的模型，构建其物理图谱和转录图谱是理解非整倍性相关表型分子基础的必要步骤。为了鉴定导致唐氏综合征先天性心脏病（DS-CHD）的基因，我们构建了从D21S55到MX1区域的物理图谱。使用跨越该区间的几个酵母人工染色体（YAC）筛选细菌人工染色体（BAC）文库，并在填补缺口的工作中使用放射性标记的序列标签位点（STS）聚合酶链反应（PCR）产物和完整的BAC筛选P1衍生人工染色体（PAC）文库。荧光原位杂交（FISH）证实了所有BAC和PAC克隆位于21q22.2-q22.3。利用克隆间Southern杂交以及从BAC和PAC末端直接测序产生的24个新STS和35个已有的STS建立重叠群。在4至5兆碱基（Mb）的从D21S55到MX1区间中，约3.5 Mb被85个BAC和24个PAC覆盖，代表重叠群内四倍覆盖。这些BAC和PAC重叠群是分离DS-CHD相关基因的宝贵试剂。
