Abstract
GM 126 human 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.
Full Text
The Full Text of this article is available as a PDF (3.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Basilico C. Temperature-sensitive mutations in animal cells. Adv Cancer Res. 1977;24:223–266. doi: 10.1016/s0065-230x(08)61016-7. [DOI] [PubMed] [Google Scholar]
- Chen T. R., McMorris F. A., Creagan R., Ricciuti F., Tischfield J., Ruddle F. Assignment of the genes for malate oxidoreductase decarboxylating to chromosome 6 and peptidase B and lactate dehydrogenase B to chromosome 12 in man. Am J Hum Genet. 1973 Mar;25(2):200–207. [PMC free article] [PubMed] [Google Scholar]
- Ming P. M., Chang H. L., Baserga R. Release by human chromosome 3 of the block at G1 of the cell cycle, in hybrids between tsAF8 hamster and human cells. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2052–2055. doi: 10.1073/pnas.73.6.2052. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nichols E. A., Elsevier S. M., Ruddle F. H. A new electrophoretic technique for mouse, human, and Chinese hamster galactokinase. Cytogenet Cell Genet. 1974;13(3):275–278. doi: 10.1159/000130279. [DOI] [PubMed] [Google Scholar]
- Nichols E. A., Ruddle F. H. A review of enzyme polymorphism, linkage and electrophoretic conditions for mouse and somatic cell hybrids in starch gels. J Histochem Cytochem. 1973 Dec;21(12):1066–1081. doi: 10.1177/21.12.1066. [DOI] [PubMed] [Google Scholar]
- Povey S., Slaughter C. A., Wilson D. E., Gormley I. P., Buckton K. E., Perry P., Bobrow M. Evidence for the assignment of the loci AK1, AK3 and ACONs to chromosome 9 in man. Ann Hum Genet. 1976 May;39(4):413–422. doi: 10.1111/j.1469-1809.1976.tb00145.x. [DOI] [PubMed] [Google Scholar]
- Sheinin R., Darragh P., Dubsky M. Mitochondrial DNA synthesis in mouse L cells temperature sensitive in nuclear DNA replication. Can J Biochem. 1977 May;55(5):543–547. doi: 10.1139/o77-077. [DOI] [PubMed] [Google Scholar]
- Sheinin R., Guttman S. Semi-conservative and non-conservative replication of DNA in temperature-sensitive mouse L-cells. Biochim Biophys Acta. 1977 Nov 2;479(1):105–118. doi: 10.1016/0005-2787(77)90130-7. [DOI] [PubMed] [Google Scholar]
- Sheinin R. Preliminary characterization of the temperature-sensitive defect in DNA replication in a mutant mouse L cell. Cell. 1976 Jan;7(1):49–57. doi: 10.1016/0092-8674(76)90254-3. [DOI] [PubMed] [Google Scholar]
- Shows T. B. Genetics of human-mouse somatic cell hybrids: linkage of human genes for isocitrate dehydrogenase and malate dehydrogenase. Biochem Genet. 1972 Dec;7(3):193–204. doi: 10.1007/BF00484817. [DOI] [PubMed] [Google Scholar]
- Slater M. L., Ozer H. L. Temperature-sensitive mutants of Balb/3T3 cells: description of a mutant affected in cellular and polyoma virus DNA synthesis. Cell. 1976 Feb;7(2):289–295. doi: 10.1016/0092-8674(76)90028-3. [DOI] [PubMed] [Google Scholar]
- Thompson L. H., Mankovitz R., Baker R. M., Till J. E., Siminovitch L., Whitmore G. F. Isolation of temperature-sensitive mutants of L-cells. Proc Natl Acad Sci U S A. 1970 Jun;66(2):377–384. doi: 10.1073/pnas.66.2.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson L. H., Mankovitz R., Baker R. M., Wright J. A., Till J. E., Siminovitch L., Whitmore G. F. Selective and nonselective isolation of temperature-sensitive mutants of mouse L-cells and their characterization. J Cell Physiol. 1971 Dec;78(3):431–440. doi: 10.1002/jcp.1040780312. [DOI] [PubMed] [Google Scholar]