Nakahori Y, Tamura T, Nagafuchi S, Fujieda K, Minowada S, Fukutani K, Fuse H, Hayashi K, Kuroki Y, Fukushima Y
Department of Congenital Abnormalities Research, National Children's Medical Research Center, Tokyo, Japan.
Genomics. 1991 Apr;9(4):765-9. doi: 10.1016/0888-7543(91)90373-m.
We have developed a novel positive cloning vector whose use precludes the cloning of any fragments less than 0.8 kb as well as 3.4-kb EcoRI fragments of DYZ1, the largest repeating-DNA family on the long arm of the human Y chromosome. Using this vector, we subcloned inserts of a Y-chromosome-specific phage library constructed from EcoRI-digested flow-sorted Y-chromosome DNA. Ten novel Y-specific fragments were obtained. Their localization on the Y chromosome was determined by deletion mapping using clinical samples with structurally abnormal Y chromosomes. The long arm of the Y chromosome was divided into 12 segments by the novel probes in combination with established probes. The amelogenin-like sequence, mapped on the long arm in Human Gene Mapping 10, has been mapped on the short arm.
我们开发了一种新型的阳性克隆载体，使用该载体可防止克隆任何小于0.8 kb的片段以及DYZ1的3.4 kb EcoRI片段，DYZ1是人类Y染色体长臂上最大的重复DNA家族。利用该载体，我们对从经EcoRI消化的流式分选Y染色体DNA构建的Y染色体特异性噬菌体文库的插入片段进行了亚克隆。获得了10个新的Y特异性片段。通过使用具有结构异常Y染色体的临床样本进行缺失定位，确定了它们在Y染色体上的定位。结合已有的探针，新型探针将Y染色体的长臂分为12个区段。在人类基因定位10中定位在长臂上的釉原蛋白样序列已被定位到短臂上。
