Cytological Mapping of Human X-Linked Genes by Use of Somatic Cell Hybrids Involving an X-Autosome Translocation

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 69; no. 1; pp. 69 - 73
• Main Authors: Grzeschik, K. H., Allderdice, P. W., Grzeschik, A., Opitz, J. M., Miller, O. J., Siniscalco, M.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01-01-1972; National Acad Sciences
• Subjects: Animals; Biological Sciences: Genetics; Cell Fusion; Cell Line; Cell lines; Chromosome Aberrations; Chromosome Mapping; Chromosome translocation; Chromosomes; Clone Cells; Cricetinae; Culture Media; Female; Genetic Linkage; Genetic loci; Glucosephosphate Dehydrogenase - metabolism; Human genetics; Human X chromosome; Humans; Hybrid Cells; Hybridity; Hypoxia; Inosine Nucleotides; Karyotyping; Mice; Mitosis; Pentosephosphates; Pentosyltransferases - metabolism; Phosphoglycerate Kinase - metabolism; Sex Chromosomes - enzymology; Somatic cells
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.69.1.69

Man-mouse and man-Syrian hamster somatic hybrid cell lines were prepared by fusion of mouse A9 or hamster TG2 cells, which are deficient in hypoxanthine-guanine phosphoribosyl transferase, with cells of a diploid fibroblastic strain, KOP-1, derived from a woman heterozygous for an X-autosome translocation. 61 clones were derived in nonselective medium and 85 sublines of these were derived in selective media: 53 in hypoxanthine-aminopterine-thymidine and 32 in 8-azaguanine. All three human X-linked markers studied, i.e., hypoxanthineguanine phosphoribosyl transferase (EC 2.4.2.8), glucose-6-phosphate dehydrogenase (EC 1.1.1.49), and phosphoglycerate kinase (EC 2.7.2.3), were present together, or absent together, in most of these clones and sublines. However, loss or retention of only phosphoglycerate kinase was occasionally observed, even in the absence of selective growth, while no evidence of separation of hypoxanthine-guanine phosphoribosyl transferase from glucose-6-phosphate dehydrogenase occurred. Cytological examination of eight man-hamster clonal lines by the quinacrine fluorescent technique showed that human phosphoglycerate kinase was only present when the translocation chromosome carrying most of the long arm of the X chromosome was present. The presence of human glucose-6-phosphate dehydrogenase and hypoxanthine-guanine phosphoribosyl transferase was not related to the presence or absence of this chromosome, but appeared to be correlated with the presence of the other translocation chromosome.