HIGH-RESOLUTION MAPPING OF HUMAN CHROMOSOME-11 BY INSITU HYBRIDIZATION WITH COSMID CLONES
HIGH-RESOLUTION MAPPING OF HUMAN CHROMOSOME-11 BY INSITU HYBRIDIZATION WITH COSMID CLONES
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DOI：
10.1126/science.2294592
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发表时间：
1990-01-05
期刊：
影响因子：
56.9
通讯作者：
WARD, DC
中科院分区：
文献类型：
--
作者：
LICHTER, P;TANG, CJC;WARD, DC
在抑制重复DNA序列信号的条件下，通过荧光原位杂交将含有人DNA插入片段的粘粒克隆定位在11号染色体上。分析了13个已知基因、1个11号染色体特异性DNA重复序列和36个随机克隆。高分辨率映射是通过使用数字成像显微镜和分析延长（前中期）染色体。通过原位杂交建立的地图坐标显示了一个一对一的对应关系，通过Southern（DNA）印迹分析的杂交细胞系含有染色体11的片段。此外，通过同时杂交三个或更多个互补序列，可以明确地建立染色体上的基因顺序。这些结果表明，通过原位杂交快速产生高分辨率的人类染色体图谱的可行性。
Cosmid clones containing human DNA inserts have been mapped on chromosome 11 by fluorescence in situ hybridization under conditions that suppress signal from repetitive DNA sequences. Thirteen known genes, one chromosome 11-specific DNA repeat, and 36 random clones were analyzed. High-resolution mapping was facilitated by using digital imaging microsocpy and by analyzing extended (prometaphase) chromosomes. The map coordinates established by in situ hybridization showed a one to one correspondence with those determined by Southern (DNA) blot analysis of hybrid cell lines containing fragments of chromosome 11. Furthermore, by hybridizing three or more cosmids simultaneously, gene order on the chromosome could be established unequivocally. These results demonstrate the feasibility of rapidly producing high-resolution maps of human chromosomes by in situ hybridization.