Lichter P, Tang C J, Call K, Hermanson G, Evans G A, Housman D, Ward D C
Department of Human Genetics, Yale University School of Medicine, New Haven, CT 06510.
Science. 1990 Jan 5;247(4938):64-9. doi: 10.1126/science.2294592.
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 microscopy 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.
在抑制重复DNA序列信号的条件下，通过荧光原位杂交技术，已将含有人DNA插入片段的黏粒克隆定位到11号染色体上。分析了13个已知基因、一个11号染色体特异性DNA重复序列和36个随机克隆。使用数字成像显微镜并分析伸展的（前中期）染色体有助于进行高分辨率定位。通过原位杂交确定的图谱坐标与通过对含11号染色体片段的杂交细胞系进行Southern（DNA）印迹分析所确定的坐标一一对应。此外，通过同时杂交三个或更多黏粒，可以明确确定染色体上的基因顺序。这些结果证明了通过原位杂交快速生成人类染色体高分辨率图谱的可行性。
