Ichikawa H, Shimizu K, Saito A, Wang D N, Oliva R, Kobayashi H, Kaneko Y, Miyoshi H, Smith C L, Cantor C R
Department of Immunology and Virology, Saitama Cancer Center Research Institute, Japan.
Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):23-7. doi: 10.1073/pnas.89.1.23.
Human chromosome 21 is the smallest of the 22 autosomes and 2 sex chromosomes. Hybridization of the human repetitive sequence Alu to pulsed-field gel-fractionated Not I-digested genomic DNA from a human-mouse hybrid cell line containing chromosome 21 as the sole human component identified chromosome 21 Not I restriction fragments. A Not I restriction map of regions of the chromosome was constructed, by identifying neighboring Alu bands with Not I linking clones. This approach simplifies the task of physical mapping and avoids ambiguities in Not I fragment assignments that arise from gel-to-gel mobility variations. A contiguous map was constructed with six Not I linking clones that covers at least the proximal one-third of the long arm of chromosome 21 and spans 20 megabases. A more detailed restriction map revealed 11 likely CpG islands in this region and localized 11 additional DNA markers.
人类21号染色体是22条常染色体和2条性染色体中最小的一条。人类重复序列Alu与来自一个人鼠杂交细胞系的脉冲场凝胶分离的Not I消化基因组DNA杂交,该细胞系以21号染色体作为唯一的人类成分,从而鉴定出21号染色体的Not I限制性片段。通过用Not I连接克隆鉴定相邻的Alu条带,构建了该染色体区域的Not I限制性图谱。这种方法简化了物理图谱绘制的任务,并避免了由于凝胶间迁移率变化而导致的Not I片段分配中的模糊性。用六个Not I连接克隆构建了一个连续图谱,该图谱至少覆盖21号染色体长臂的近端三分之一,跨度为20兆碱基。一个更详细的限制性图谱揭示了该区域有11个可能的CpG岛,并定位了另外11个DNA标记。