Tanahashi H
Laboratory of Molecular Medicine, University of Tokyo.
Nihon Rinsho. 1994 Feb;52(2):515-25.
To clarify the structure of human chromosome 21q, we isolated many Sequence-Tagged NotI and SfiI linking clones. For further analysis, we constructed a P1 phage library specific for this chromosome. The library presently consists of 1516 clones, amounting to 2 equivalents of this chromosome. For rapid clone identification, we constructed a 3-dimensional library containing DNA from P1 clones, and a PCR-based screening approach was used to isolate 38 positives from 50 sequence-tagged sites specific for this chromosome. And also by this PCR-based chromosome walking procedure using STSs from sequences from insert-end of P1 clones, we constructed a contig spanning the entirety of the human beta-amyloid precursor protein gene consisted of eight P1 clones and a contig consisted of nine P1 clones on D21S8 region.
为了阐明人类21号染色体q臂的结构，我们分离了许多序列标签NotI和SfiI连接克隆。为了进一步分析，我们构建了一个针对该染色体的P1噬菌体文库。该文库目前由1516个克隆组成，相当于该染色体的2倍量。为了快速鉴定克隆，我们构建了一个包含来自P1克隆DNA的三维文库，并采用基于PCR的筛选方法从50个该染色体特异性序列标签位点中分离出38个阳性克隆。此外，通过使用来自P1克隆插入末端序列的STS进行基于PCR的染色体步移程序，我们构建了一个跨越整个人类β-淀粉样前体蛋白基因的重叠群，该重叠群由8个P1克隆组成，以及一个在D21S8区域由9个P1克隆组成的重叠群。
