Analysis of the 1.1-Mb human alpha/beta T-cell receptor locus with bacterial artificial chromosome clones
Analysis of the 1.1-Mb human alpha/beta T-cell receptor locus with bacterial artificial chromosome clones
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DOI：
10.1101/gr.7.4.330
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发表时间：
1997-04-01
期刊：
影响因子：
7
通讯作者：
Hood, L
中科院分区：
文献类型：
--
作者：
Boysen, C;Simon, MI;Hood, L
细菌人工染色体（BAC）克隆是有效的作图和测序试剂。人类1.1-Mb α/δ T细胞受体基因座的定位和BAC克隆的部分测序。17个BAC克隆覆盖了1.1-Mb α/δ基因座，除了一个小的缺口，该缺口是从3.7倍BAC文库可能提供的覆盖度预期的。BAC插入片段的末端序列可以通过链终止子化学测序直接从BAC DNA获得。五个完整的BAC插入直接测序的鸟枪法。17个BAC插入物的末端均匀分布在整个基因座上。通过几个独立的标准，BAC克隆忠实地代表了基因组DNA，除了一个68 kb缺失的克隆。这些BAC特征导致了一种新的人类基因组测序方法的提出。
Bacterial artificial chromosome (BAC) clones are effective mapping and sequencing reagents. The 1.1-Mb alpha/delta T-cell receptor locus of humans was mapped and partially sequenced with BAC clones. Seventeen BAC clones covered the 1.1-Mb alpha/delta locus, with the exception of one small gap that was expected from the coverage that a 3.7-foId BAC library is likely to provide. The end sequences of the BAC inserts could be obtained directly from the BAC DNA by sequencing with the chain terminator chemistry. Five complete BAC inserts were sequenced directly by the shotgun approach. The ends of the 17 BAC inserts were distributed evenly across the locus. By several independent criteria, the BAC clones faithfully represented the genomic DNA, with the exception of a single clone with a 68-kb deletion. These BAC features led to the proposal of a new approach to sequence the human genome.