Integration of a dominant selectable marker into human chromosomes and transfer of marked chromosomes to mouse cells by microcell fusion.
A method for the production of stable mouse-human cell hybrids containing a single human chromosome is described. As a first step in this method, a cloned selectable marker, the E. coli xanthine-guanine phosphoribosyltransferase (Ecogpt) gene, was transferred to human cells to generate cell lines each carrying Ecogpt integrated into a different site. Human chromosomes marked with Ecogpt were transferred further into mouse cells by microcell fusion. Monochromosomal hybrids, in which the human chromosome is maintained by selection, have been produced for chromosomes 2, 5, 16, and a rearranged chromosome involving a translocation between chromosomes 1 and 2. In addition to these monochromosomal hybrids, we have also obtained monochromosomal hybrids for human chromosomes 6, 12, and 17 by selection for the loss of marked chromosome from the microcell hybrids each containing two human chromosomes. Although the human chromosome present in these hybrids cannot be maintained by selection, 80-90% of cells retained the transferred chromosome on continuous growth for 15 days. Monochromosomal hybrids would provide biological materials to construct genetic maps of human chromosomes. In addition, chromosomes marked with dominant selectable markers can be transferred further to any cell line of interest in inter- or intra-species combination.
D000818:Animals; D002459:Cell Fusion; D002460:Cell Line; D002877:Chromosomes, Human; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells; D007527:Isoenzymes / Q000235:genetics; D007621:Karyotyping; D051379:Mice; D009693:Nucleic Acid Hybridization; D010430:Pentosyltransferases / Q000235:genetics; D014162:Transfection
D004269:DNA, Bacterial; D005819:Genetic Markers; D007527:Isoenzymes; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase