Genetic and physical mapping of the Lps locus:: Identification of the Toll-4 receptor as a candidate gene in the critical region
Genetic and physical mapping of the Lps locus:: Identification of the Toll-4 receptor as a candidate gene in the critical region
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DOI：
10.1006/bcmd.1998.0201
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发表时间：
1998-09-15
影响因子：
2.3
通讯作者：
Beutler, B
中科院分区：
文献类型：
--
作者：
Poltorak, A;Smirnova, I;Beutler, B
基于在两个单独的种内回交中分析的2093个减数分裂，小鼠Lps（d）突变的位置被限制在大小为0.9cM的遗传间隔内。共19个遗传标记，位于靠近突变进行了检查，在映射。其中大部分是以前未发表的多态性微卫星，通过跨越感兴趣区域的YAC和BAC克隆的片段化鉴定。发现Lps（d）与微卫星标记D4 MIT 178和在D4 MIT 178附近鉴定的三个新的多态性微卫星不可分。“B位于突变的着丝粒，在一组1600只小鼠中被4个交叉分开; 83.3位于突变的远端，在一组493只小鼠中被3个交叉分开。66个BAC克隆和1个YAC克隆被组装到临界区的95%以上。基于脉冲场凝胶电泳和荧光原位杂交的估计表明，B --> 83.3区间的长度约为3.2 M B。还指定了与Lps（d）零重组距离的最小区域，发现其占据约1.2 Mb的物理大小。为了识别候选基因，在关键区域进行了近40，000次测序。还采用选择性杂交和外显子捕获来鉴定整个“零”区域的基因。在整个关键区域内仅鉴定出单个完整基因。该基因编码Toll-4受体，其为IL-1受体家族的成员，（C）1998 Academic Press
On the basis of 2093 meioses analyzed in two separate intraspecific backcrosses, the location of the mouse Lps(d) mutation was circumscribed to a genetic interval 0.9 cM in size. A total of 19 genetic markers that lie in close proximity to the mutation were examined in mapping. Most of these were previously unpublished polymorphic microsatellites, identified by fragmentation of YAC and BAC clones spanning the region of interest, Lps(d) was found to be inseparable from the microsatellite marker D4MIT178, and from three novel polymorphic microsatellites identified near D4MIT178, The mutation was confined between two novel microsatellite markers, herein designated "B" and "83.3." B lies centromeric to the mutation, and was separated by four crossovers in a panel of 1600 mice; 83.3 lies distal to the mutation and was separated by three crossovers in a panel of 493 mice. 66 BAC clones and one YAC clone were assembled to cover >95% of the critical region, Estimates based on pulsed field gel electrophoresis and fluorescence in situ hybridization indicate that the The B --> 83.3 interval is about 3.2 Mb in length. An minimal area of zero recombinational distance from Lps(d) was also assigned, and found to occupy approximately 1.2 Mb of physical size. To identify gene candidates, nearly 40,000 sequencing runs were performed across the critical region. Selective hybridization and exon trapping were also employed to identify genes throughout the "zero" region. Only a single intact gene was identified within the entire critical region. This gene encodes the Toll-4 receptor, a member of the IL-1 receptor family, (C) 1998 Academic Press