Jeggo P A, Hafezparast M, Thompson A F, Kaur G P, Sandhu A K, Athwal R S
MRC Cell Mutation Unit, Sussex University, Falmer, Brighton, U.K.
Somat Cell Mol Genet. 1993 Jan;19(1):39-49. doi: 10.1007/BF01233953.
We have constructed hamster-human hybrid cell lines containing fragments of human chromosome 2 as their only source of human DNA. Microcell-mediated chromosome transfer was used to transfer human chromosome 2 from a monochromosomal mouse-human hybrid line to a radiation-sensitive hamster mutant (XR-V15B) defective in double-strand break rejoining. The human chromosome 2 carried the Ecogpt gene and hybrids were selected using this marker. The transferred human chromosome was frequently broken, and the resulting microcell hybrids contained different sized segments of the q arm of chromosome 2. Two microcell hybrids were irradiated and fused to XR-V15B to generate additional hybrids bearing reduced amounts of human DNA. All hybrids were analyzed by PCR using primers specific for 27 human genes located on chromosome 2. From these data we have localized the integrated gpt gene on the human chromosome 2 to the region q36-37 and present a gene order for chromosome 2 markers.
我们构建了仓鼠-人类杂交细胞系,其中包含人类2号染色体片段作为其唯一的人类DNA来源。利用微细胞介导的染色体转移技术,将人类2号染色体从一个单染色体的小鼠-人类杂交系转移到一个在双链断裂修复方面存在缺陷的辐射敏感仓鼠突变体(XR-V15B)中。人类2号染色体携带Ecogpt基因,并使用该标记筛选杂种细胞。转移的人类染色体经常发生断裂,产生的微细胞杂种包含2号染色体q臂不同大小的片段。对两个微细胞杂种进行辐射并与XR-V15B融合,以产生携带更少人类DNA量的额外杂种细胞。使用针对位于2号染色体上的27个人类基因的特异性引物,通过PCR对所有杂种细胞进行分析。根据这些数据,我们已将整合在人类2号染色体上的gpt基因定位到q36-37区域,并给出了2号染色体标记的基因顺序。