CONSTRUCTION OF A GENERAL HUMAN-CHROMOSOME JUMPING LIBRARY, WITH APPLICATION TO CYSTIC-FIBROSIS
CONSTRUCTION OF A GENERAL HUMAN-CHROMOSOME JUMPING LIBRARY, WITH APPLICATION TO CYSTIC-FIBROSIS
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DOI：
10.1126/science.2950591
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发表时间：
1987-02-27
期刊：
影响因子：
56.9
通讯作者：
IANNUZZI, MC
中科院分区：
文献类型：
--
作者：
COLLINS, FS;DRUMM, ML;IANNUZZI, MC
在许多遗传性疾病中，负责的基因及其蛋白质产物是未知的。这种技术被称为“反向遗传学”，其中染色体图谱位置和遗传连锁的 DNA 标记用于识别和克隆此类基因，但由于从最近的 DNA 标记到基因本身的分子距离通常太大而无法通过标准克隆技术遍历，因此该技术变得复杂。为了解决这种情况，构建了一个通用的人类染色体跳跃文库，该文库允许克隆距离基因组 DNA 中任何起始点约 100 KB 的 DNA 序列。为了说明其有用性，我们从met癌基因开始搜索该文库寻找跳跃克隆，met癌基因是与位于人类7号染色体上的囊性纤维化基因紧密连锁的标记物。通过脉冲场凝胶电泳对新基因组片段进行定位，证实它位于7号染色体上met基因下游240 kilobase内。染色体跳跃的使用现在应该适用于任何有紧密连锁 DNA 标记的遗传位点。
In many genetic disorders, the responsible gene and its protein product are unknown. The technique know as "reverse genetics," in which chromosomal map positions and genetically linked DNA markers are used to identify and clone such genes, is complicated by the fact that the molecular distances from the closest DNA markers to the gene itself are ofter too large to traverse by standard cloning techniques. To address this situation, a general human chromosome jumping library was constructed that allows the cloning of DNA sequences approximately 100 kilobases away from any starting point in genomic DNA. As an illustration of its usefulness, this library was searched for a jumping clone, starting at the met oncogen, which is a marker tighly linked to the cystic fibrosis gene that is located on human chromosome 7. Mapping of the new genomic fragment by pulsed field gel electrophoresis confirmed that it resides on chromosome 7 within 240 kilobases downstream of the met gene. The use of chromosome jumping should now be applicable to any genetic locus for which a closely linked DNA marker is available.