Bickmore W A, Maule J C, van Heyningen V, Porteous D J
MRC Human Genetics Unit, Western General Hospital, Edinburgh, Scotland.
Somat Cell Mol Genet. 1989 May;15(3):229-35. doi: 10.1007/BF01534873.
We have used chromosome-mediated gene transfer (CMGT) and whole cell fusion to derive human-mouse hybrid cells carrying reduced human chromosomes 11, by selecting for expression of the transforming H-ras 1 oncogene. To realize the full potential of these somatic cell genetic techniques as resources for enriched DNA probe isolation and the fine structure mapping of chromosomes, the nature of any molecular rearrangements that may accompany the process of DNA transfer must be understood. We have analyzed the long-range structure of our transgenomes by pulsed field gel electrophoresis (PFGE) and show here that, whereas during cell fusion several megabase pairs (Mb) of DNA can be transferred intact, multiple rearrangements of DNA accompany CMGT even in transgenomes where other methods of analysis gave no indication of such molecular scrambling.
我们利用染色体介导的基因转移(CMGT)和全细胞融合技术,通过选择转化型H-ras 1癌基因的表达,获得了携带人类11号染色体片段减少的人-鼠杂交细胞。为了充分发挥这些体细胞遗传技术作为富集DNA探针分离和染色体精细结构图谱绘制资源的潜力,必须了解DNA转移过程中可能伴随的任何分子重排的性质。我们通过脉冲场凝胶电泳(PFGE)分析了转基因组的长程结构,结果表明,在细胞融合过程中,几兆碱基对(Mb)的DNA可以完整转移,而在CMGT过程中,即使在其他分析方法未显示这种分子混乱迹象的转基因组中,DNA也会发生多次重排。