SEGREGATION OF THE HUNTINGTON DISEASE REGION OF HUMAN-CHROMOSOME 4 IN A SOMATIC-CELL HYBRID
SEGREGATION OF THE HUNTINGTON DISEASE REGION OF HUMAN-CHROMOSOME 4 IN A SOMATIC-CELL HYBRID
复制标题
DOI：
10.1016/0888-7543(89)90347-9
复制
发表时间：
1989-04-01
期刊：
影响因子：
4.4
通讯作者：
MYERS, RM
中科院分区：
文献类型：
--
作者：
COX, DR;PRITCHARD, CA;MYERS, RM
我们已经开发了一种x射线照射细胞融合程序，分离缺乏可选择标记的人类染色体片段，并使用这种方法构建保留人类4号染色体片段和唯一人类材料的体细胞杂交。为了鉴定在亨廷顿病（HD）基因区域保留小染色体片段的杂交种，我们使用Southern blot分析筛选了72个杂交种，以确定7个4号染色体单拷贝位点的存在或缺失。这些数据与原位杂交实验相结合，确定了三种感兴趣的杂交。其中一种细胞系C25在HD基因附近稳定地保留了1万到2万kb的远端4p片段，并易位到仓鼠染色体上。场反转凝胶电泳未发现C25中存在人类DNA重排的证据。结合类似的辐射杂交，C25是分离HD基因附近DNA探针的有价值的工具。
We have developed an X-irradiation:cell fusion procedure that segregate segments of human chromosomes lacking selectable markers and have used this approach to construct somatic cell hybrids retaining fragments of human chromosome 4 and the only human material. To identify hybrids retaining a small chromosomal fragment in the region of the Huntington disease (HD) gene, we used Southern blot anlaysis to screen 72 hybrid lines for the presence or absence of seven chromosome 4 single-copy loci. These data combined with in situ hybridization experiments, identified three hybrids of interest. One of these cell lines C25, stably retains a 10,000- to 20,000-kb fragment of distal 4p in the vicinity of the HD gene, translocated to a hamster chromosome. Field-inversion gel electrophoresis revealed no evidence of rearrangements in the human DNA present in C25. In combination with similar radiation hybrids, C25 is a valuable tool for isolating DNA probes near the HD gene.