X-LINKAGE OF A HUMAN GENETIC LOCUS THAT CORRECTS THE DNA SYNTHESIS LESION IN tsC1AGOH MOUSE CELLS

作者：Richard E. Giles, Frank H. Ruddle · 发表于：Genetics · 年份：1979 · DOI：10.1093/genetics/93.4.975 · 被引用次数：12 · 研究领域：CRISPR and Genetic Engineering、Animal Genetics and Reproduction、Pluripotent Stem Cells Research

GM 126 diploid fibroblasts were fused with a heat-sensitive mouse cell mutant defective in DNA synthesis, and primary hybrids were selected at permissive and nonpermissive temperatures in HAT medium. Primary hybrids, primary hybrid clones back-selected in 8-azaguanine at the permissive temperature, and subclones of heat-resistant primary hybrids isolated under nonselective conditions or after 8-azaguanine treatment were tested for heat sensitivity, the expression of 26 human enzymes assigned to 19 different human chromosomes, and the presence of human chromosomes. Only the human X chromosome and X-linked marker enzymes exhibited a clear pattern of concordant segregation with the heat-resistant phenotype. On the basis of these observations, we have defined the human genetic locus that corrects the heat-sensitive lesion in tsC1AGOH as hrC1AGOH and have assigned this locus to the X chromosome. This observation provides the first instance where two selectable markers (heat resistance and 8-azaguanine sensitivity) are found on a single human chromosome and suggests that these markers may prove to be a valuable push-pull selective system of use in determining the linear arrangement of genes on human chromosomes by somatic cell genetics.