Mapping of human chromosome 11: organization of genes within the Wilms' tumor region of the chromosome.
Mapping of human chromosome 11: organization of genes within the Wilms' tumor region of the chromosome.
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人类 11 号染色体作图：染色体肾母细胞瘤区域内基因的组织。
DOI：
10.1101/sqb.1986.051.01.097
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Lewis,WH
中科院分区：
文献类型：
--
作者：
Housman,DE;Glaser,T;Gerhard,DS;Jones,C;Bruns,GA;Lewis,WH
结果为了准确定位Wilms‘s肿瘤基因，我们对WAGR患者11号染色体缺失的DNA序列进行了分析。在一组重叠的此类缺失中定位DNA序列将为最终分离WAGR复合体内的基因提供必要的重要信息。在这项分析中遇到的一个技术问题是，携带这种缺失的个体的细胞中存在正常的11号染色体同源基因。WAGR患者的DNA中存在两个11号染色体同源物，这使得除非进行定量的基因剂量研究，否则很难分析DNA片段的副本是否被删除。为了分离单独的11号染色体同源物，我们将WAGR患者的体细胞与已建立的啮齿动物细胞系融合。通过这种方式，我们可以通过杂交细胞系基因组DNA的Southern分析直接确定给定的DNA序列是否存在于WAGR区域内。图1显示了两名携带WAGR缺失的患者的分析实例。在本实验中，
RESULTSTo map the Wilms' tumor gene precisely, we have characterized DNA sequences deleted in the chromosomes 11 of WAGR patients. Locating DNA sequences within an overlapping set of such deletions will provide important information necessary for the ultimate isolation of the genes within the WAGR complex. A technical problem encountered in this analysis is the presence of the normal chromosome 11 homolog in the cells of individuals carrying such deletions. The presence of two chromosome 11 homologs in DNA from WAGR patients makes it difficult to analyze whether a copy of a DNA segment is deleted unless quantitative gene dosage studies are performed. To separate the individual chromosome 11 homologs, we fused the somatic cells of WAGR patients with established rodent cell lines. In this manner we could determine directly whether a given DNA sequence was present or absent within the WAGR region by Southern analysis of genomic DNA from hybrid cell lines. An example of this analysis for two patients carrying WAGR deletions is shown in Figure 1. In this experi-