Hirotsune S, Takahara T, Sasaki N, Imoto H, Okazaki Y, Eki T, Murakami Y, Abe M, Furuya K, Muramatsu M, Eto Y, Chapman V M, Hayashizaki Y
RIKEN Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
Genomics. 1996 Oct 1;37(1):87-95. doi: 10.1006/geno.1996.0524.
We have established a new system for chromosome-specific yeast artificial chromosome (YAC) contig construction using restriction landmark genomic scanning (RLGS-based YAC contig mapper). RLGS is a powerful tool for detecting more than 1000 restriction landmarks distributed on an entire genome in one procedure. In this system, RLGS is applied to sorted chromosomes to cover the target chromosome. Using these landmarks as guideposts, chromosome-specific YAC clones are then ordered. In this paper, we report the construction of a map for a human chromosome 21 YAC contig spanning q22.1 using this new approach. Applying RLGS to sorted chromosomes 21 enables detection of approximately 1400 spots (equivalent of 1050 PacI landmarks), covering the entire region of this chromosome. We constructed the 2.5-Mb YAC contig encompassing 21q22.1 with 66 spots (equivalent of 50 PacI landmarks). With this contig map, we could detect two deleted regions and chimerism in the YAC insert DNA. Our results demonstrated the usefulness of this approach for finding DNA alterations of YACs, such as deletions and chimerism.
我们已经建立了一种新的系统，用于构建特定染色体的酵母人工染色体（YAC）重叠群，该系统使用限制酶切位点基因组扫描技术（基于RLGS的YAC重叠群映射器）。RLGS是一种强大的工具，可在一个步骤中检测分布在整个基因组上的1000多个限制酶切位点。在该系统中，RLGS应用于分选后的染色体以覆盖目标染色体。然后，以这些酶切位点为路标，对特定染色体的YAC克隆进行排序。在本文中，我们报告了使用这种新方法构建跨越21q22.1的人类21号染色体YAC重叠群图谱的过程。将RLGS应用于分选后的21号染色体，能够检测到大约1400个位点（相当于1050个PacI酶切位点），覆盖该染色体的整个区域。我们构建了包含21q22.1的2.5 Mb YAC重叠群，其中有66个位点（相当于50个PacI酶切位点）。利用这个重叠群图谱，我们能够检测到YAC插入DNA中的两个缺失区域和嵌合现象。我们的结果证明了这种方法在发现YAC的DNA改变（如缺失和嵌合现象）方面的有效性。
