Characterisation of molecular DNA rearrangements within the Xq12-q13.1 region, in three patients with X-linked hypohidrotic ectodermal dysplasia (EDA).
Characterisation of molecular DNA rearrangements within the Xq12-q13.1 region, in three patients with X-linked hypohidrotic ectodermal dysplasia (EDA).
复制标题
三名 X 连锁少汗性外胚层发育不良 (EDA) 患者 Xq12-q13.1 区域内分子 DNA 重排的特征。
DOI：
10.1093/hmg/2.10.1679
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发表时间：
1993
影响因子：
3.5
通讯作者：
Clarke,A
中科院分区：
文献类型：
--
作者：
Thomas,NS;Chelly,J;Zonana,J;Davies,KJ;Morgan,S;Gault,J;Rack,KA;Buckle,VJ;Brockdorff,N;Clarke,A
一组体细胞杂交和X连锁少汗性外胚叶发育不良(EDA)患者来源的细胞系，包含不同重排的X染色体，已被用于完善Xq12-q13.1区域的物理图谱。患者来源的材料包括来自一个间质缺失的EDA男性(EDA家族1015)的基因组DNA，从一个孤立的(X；9)易位的女性患者获得的细胞系GM0705A，以及来自该细胞系的体细胞杂种Anly。该地图将该区域细分为至少6个映射间隔。利用DXS732和DXS453的DNA探针对酵母人工染色体(YAC)克隆进行了鉴定。荧光原位杂交(FISH)分析显示了两个DXS732特异的YAC，以桥接(X；9)易位断裂点。这两个YAC还与ICRF人类X染色体粘粒文库进行了筛选，鉴定了36个粘粒克隆。直接粘粒-粘粒杂交分析将这些克隆的子集放置在四个不同的粘粒重叠群中。对照映射面板，映射来自每个重叠群的锚定克隆，将所有这些重叠群定位在Xql2-Qi3.1区域内。一个粘粒，ICRFc104C03.1 84，在Anly杂交DNA的几个限制性内切酶中发现了潜在的连接片段。对具有总粘粒ICRFc104C03.184的GMO705A细胞系的FISH分析证实了这一点，在该细胞系中鉴定出了(X；9)易位的两个染色体元件。来自这个宇宙的单拷贝探针pC03.184F5在与Anly DNA杂交时也发现了Der(9)衍生的连接片段。从80个受pC03.184F5影响的无关EDA男性中筛选大量基因组DNA，除了最初的EDA 1015缺失外，在第二个不相关的EDA男性(EDA家族1003)中发现了一个分子缺失。这两个EDA缺失的特征表明，EDA 1003缺失的近端终点位于DXS452基因座和EDA 1015缺失的近端终点的远端。EDA 1015缺失的远端终点位于四个粘粒重叠群的最远端。EDA 1003缺失的远端终点仍有待确定，但必须位于DXS453基因座的近端。确定EDA 1003缺失的程度和进一步分析(X；9)易位的粘粒，应该会极大地促进EDA基因的定位克隆。
A panel of somatic cell hybrids and X-linked hypohidrotic ectodermal dysplasia (EDA) patient-derived cell lines, containing different rearranged X chromosomes, have been used to refine the physical map of the Xq12–q13.1 region. The patient-derived material included genomic DNA from an EDA male (EDA family 1015) with an interstitial deletion , and a cell line GM0705A, obtained from an isolated female patient with ade novobalanced (X;9) translocation, and the somatic hybrid, AnLy, derived from this cell line. This map subdivides the region into at least 6 mapping-intervals. DNA probes from DXS732 and DXS453, identified as the closest flanking marker loci to the EDA locus, were used to identify homologous Yeast Artificial Chromosome (YAC) clones. Two of the DXS732-specific YACs were shown by fluorescentin situhybridisation (FISH) analysis to bridge the (X;9) translocation breakpoint. These two YACs were also screened against the ICRF human X chromosome cosmid library and identified 36 cosmid clones. Direct cosmid-cosmid hybridisation analysis placed subsets of these clones within four different cosmid contigs. Mapping of anchor clones from each contig, against the mapping panel, localised all these contigs within the Xql2–qi3.1 region. One cosmid, ICRFc104C03.1 84, identified potential junctional-fragments in several restriction digests of AnLy hybrid DNA. This was confirmed by FISH analysis of the GMO705A cell line with total cosmid ICRFc104C03.184, In which both chromosomal elements of the (X;9) translocation were identified. A single-copy probe pC03.184F5, derived from this cosmld, also Identified the der(9)- derived junctional fragment when hybridised against AnLy DNA. Screening a large panel of genomic DNAs from 80 unrelated affected EDA males with pC03.184F5 identified, in addition to the original EDA 1015 deletion, a molecular deletion in a second unrelated EDA male (EDA family 1003). Characterisation of these two EDA deletions has shown that the proximal endpoint of the EDA 1003 deletion lies distal, to both the DXS452 locus and to the proximal endpoint of the EDA 1015 deletion. The distal endpoint of the EDA 1015 deletion has been located within the most distal of the four cosmid contigs. The distal endpoint of the EDA 1003 deletion has still to be established, but must be located proximal of the DXS453 locus. Determination of the extent of the EDA 1003 deletion and further analysis of the (X;9) translocation-fianking cosmids, should greatly facilitate the positional cloning of the EDA gene.