Pritchard C, Goodfellow P N
EMBO J. 1986 May;5(5):979-85. doi: 10.1002/j.1460-2075.1986.tb04312.x.
Chromosome-mediated gene transfer (CMGT) can be used to generate fragments of human chromosomes and chromosomal maps can be constructed using these fragments. In previous experiments CMGT techniques have been limited to those regions of the genome which encode biochemically selectable markers. We have extended the regions of the human genome which can be subjected to CMGT methods by employing a cell surface antigen as a selectable marker. These experiments have been facilitated by the discovery that co-transformation of chromosomes with a plasmid bearing a biochemically selectable marker followed by selection for the marker pre-selects for cells which have incorporated chromosomal fragments. The plasmid may also integrate into the donor chromosomes and this provides, in some cases, an additional selectable marker in the chromosome fragment of interest. Using these methods we have isolated for the first time cells containing varying portions of the human Y chromosome.
染色体介导的基因转移(CMGT)可用于生成人类染色体片段,并且可以使用这些片段构建染色体图谱。在之前的实验中,CMGT技术仅限于基因组中编码生物化学可选择标记的区域。我们通过使用细胞表面抗原作为选择标记,扩展了可应用CMGT方法的人类基因组区域。染色体与携带生物化学可选择标记的质粒共转化,然后选择该标记,这一发现促进了这些实验,因为它预先选择了已整合染色体片段的细胞。质粒也可能整合到供体染色体中,在某些情况下,这为感兴趣的染色体片段提供了额外的选择标记。使用这些方法,我们首次分离出了含有不同部分人类Y染色体的细胞。