Law M L, Davidson J N, Kao F T
Proc Natl Acad Sci U S A. 1982 Dec;79(23):7390-4. doi: 10.1073/pnas.79.23.7390.
Recombinant lambda phage Charon 4A with repetitive human DNA inserts have been constructed by using cellular DNA from a human-Chinese hamster ovary cell hybrid retaining the complete hamster genome and a single human chromosome 12. One recombinant phage, 12-11, contains several repetitive sequences, each with a different repetition pattern in the human genome. A 2.2-kilobase (kb) EcoRI fragment of this phage was subcloned in pBR325. This sequence has fewer than 5,000 copies in the human genome and does not cross-hybridize with Chinese hamster DNA. When the labeled 2.2-kb probe was hybridized to human chromosome 12 DNA digested with EcoRI, there was an intense band at the 2.2-kb position and a series of other discrete bands. The band pattern at positions other than 2.2 kb appears to be distinct for each human chromosome. The 2.2-kb fragment is composed of at least three subregions. The ends of the fragment are repeated more frequently in the genome than is the middle portion. Hybridization of chromosome 12 DNA with probes made to these subregions yielded simpler band patterns. By using a series of cell hybrids containing various deletions of human chromosome 12, five sequences related to the 2.2-kb fragment have been assigned regionally to a specific portion of the short arm of chromosome 12. These results demonstrate that certain repetitive sequences in the human genome can be used as genetic markers and may permit detailed regional mapping of human chromosomes.
通过使用来自保留完整仓鼠基因组和一条人类12号染色体的人-中国仓鼠卵巢细胞杂种的细胞DNA，构建了带有重复人类DNA插入片段的重组λ噬菌体Charon 4A。一种重组噬菌体12 - 11含有几个重复序列，每个序列在人类基因组中具有不同的重复模式。该噬菌体的一个2.2千碱基（kb）的EcoRI片段被亚克隆到pBR325中。这个序列在人类基因组中的拷贝数少于5000个，并且不与中国仓鼠DNA交叉杂交。当用标记的2.2 - kb探针与经EcoRI消化的人类12号染色体DNA杂交时，在2.2 - kb位置出现一条强带以及一系列其他离散带。在2.2 kb以外位置的带型似乎对于每个人类染色体都是独特的。2.2 - kb片段由至少三个亚区域组成。该片段的末端在基因组中的重复频率高于中间部分。用针对这些亚区域制备的探针与12号染色体DNA杂交产生了更简单的带型。通过使用一系列包含人类12号染色体各种缺失的细胞杂种，与2.2 - kb片段相关的五个序列已被区域定位到12号染色体短臂的特定部分。这些结果表明，人类基因组中的某些重复序列可作为遗传标记，并可能允许对人类染色体进行详细的区域图谱绘制。
