Isolation of cDNAs from the Cri-du-chat critical region by direct screening of a chromosome 5-specific cDNA library.
Isolation of cDNAs from the Cri-du-chat critical region by direct screening of a chromosome 5-specific cDNA library.
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DOI：
10.1101/gr.7.2.118
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发表时间：
1997-02
期刊：
影响因子：
7
通讯作者：
A. Simmons;J. Overhauser;M. Lovett
中科院分区：
文献类型：
--
作者：
A. Simmons;J. Overhauser;M. Lovett
染色体特异性cDNA文库是从特定基因组区域分离基因的新工具。我们使用两个YAC跨越染色体5 p的约2 Mb的cri-du-chat关键区（CDCCR）直接筛选染色体5特异性（CH 5SP）胎脑cDNA文库。为了将该文库与用于新基因发现的其他来源进行比较，将YAC与已广泛用于表达序列标签（EST）生成的标准化婴儿脑（NIB）cDNA文库杂交。这些筛选从CH 5SP胎脑文库中产生12个cDNA，从NIB文库中产生4个cDNA，其映射到CDCCR内的离散间隔。四个cDNA映射在最小的CDCCR删除间隔内，其余的cDNA位于边界之外。在CH 5SP和NIB克隆组之间只有一个cDNA共享序列重叠。其余11个CH 5SP cDNA与EST序列均不同源，这表明，与这些文库的先前数据相同，染色体特异性cDNA文库是新表达序列的丰富来源。与NIB文库重叠的单个cDNA包含与血小板反应蛋白、备解素和几种补体蛋白共享的序列基序的两个拷贝。该基序通常存在于粘附蛋白中，并且似乎介导细胞-细胞或细胞-底物相互作用。这个新的血小板反应蛋白样基因，以及其他三个在CDCCR中定位的cDNA，代表了cri-du-chat连续基因缺失综合征的候选基因。
Chromosome-specific cDNA libraries are new tools for the isolation of genes from specific genomic regions. We have used two YACs than span the approximately 2-Mb cri-du-chat critical region (CDCCR) of chromosome 5p to directly screen a chromosome 5-specific (CH5SP) fetal brain cDNA library. To compare this library with other sources for new gene discovery, the YACs were hybridized to a normalized infant brain (NIB) cDNA library that has been used extensively for expressed sequence tag (EST) generation. These screens yielded 12 cDNAs from the CH5SP fetal brain library and four cDNAs from the NIB library that mapped to discrete intervals within the CDCCR. Four cDNAs mapped within the minimal CDCCR deletion interval, with the remaining cDNAs being located beyond the boundaries. Only one cDNA shared sequence overlap between the CH5SP and NIB sets of clones. None of the remaining 11 CH5SP cDNAs were homologous to EST sequences, suggesting, in common with previous data on these libraries, that chromosome-specific cDNA libraries are a rich source of new expressed sequences. The single cDNA that did overlap with the NIB library contained two copies of a sequence motif shared with thrombospondin, properdin, and several complement proteins. This motif is usually present in adhesive proteins, and appears to mediate cell-cell or cell-substrate interactions. This new thrombospondin-like gene, and the other three cDNAs that map within the CDCCR, represent candidate genes for the cri-du-chat contiguous gene deletion syndrome.