Shortle V P, Malaspina P, Fox M, Dooley J S, Volz A, Ziegler A, Trowsdale J, Morrison K M, Edwards Y H
Academic Department of Medicine, Royal Free Hospital, School of Medicine, London, UK.
Mamm Genome. 1993 Sep;4(9):493-8. doi: 10.1007/BF00364783.
We have explored the potential of irradiation-fusion gene transfer (IFGT) hybrids as a source of well-defined human chromosome fragments from which probes can be derived. Extensive characterization of the IFGT hybrid 4J4 with a full panel of markers from Chromosome (Chr) 6 showed that the human DNA content derives largely from 6p21.3 and 6q27. A cosmid library has been constructed from 4J4 DNA, and 370 recombinants containing human DNA have been isolated and overlapping clones ordered into 20 contigs. Regional localization of representative clones from each contig, determined by fluorescent in situ hybridization (FISH), places 13 contigs in 6q27 and 6 in 6p21.3. Preliminary screening of cDNA libraries with selected cosmids has identified two expressed sequences. Since there are a number of medically important genes in both these regions of human Chr 6 with several disease loci linked to the HLA-A region in 6p21.3 and various tumor suppressor genes to 6q27, this library will provide a valuable resource to aid the isolation of candidate genes for these diseases. In addition, unique markers for detailed physical and genetic mapping of these regions of human Chr 6 can be easily obtained.
我们已经探索了辐射融合基因转移（IFGT）杂种作为明确的人类染色体片段来源的潜力，从中可以衍生出探针。用来自6号染色体（Chr）的全套标记对IFGT杂种4J4进行了广泛表征，结果表明人类DNA含量主要来自6p21.3和6q27。已从4J4 DNA构建了一个黏粒文库，分离出370个含有人DNA的重组体，并将重叠克隆排列成20个重叠群。通过荧光原位杂交（FISH）确定每个重叠群中代表性克隆的区域定位，将13个重叠群定位在6q27，6个定位在6p21.3。用选定的黏粒对cDNA文库进行初步筛选，已鉴定出两个表达序列。由于人类6号染色体的这两个区域都有许多医学上重要的基因，6p21.3中有几个疾病位点与HLA - A区域相关，6q27中有各种肿瘤抑制基因，因此该文库将为分离这些疾病的候选基因提供宝贵资源。此外，还可以轻松获得用于人类6号染色体这些区域详细物理和遗传图谱绘制的独特标记。
