Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1971 Jan;68(1):82–86. doi: 10.1073/pnas.68.1.82

A New Reduced Human-Mouse Somatic Cell Hybrid Containing the Human Gene for Adenine Phosphoribosyltransferase

Toshihisa Kusano 1,*, Cedric Long 1,†, Howard Green 1,‡
PMCID: PMC391107  PMID: 5276304

Abstract

A system that selects for the gene directing synthesis of the enzyme adenine phosphoribosyltransferase (APRT) uses the antibiotic alanosine to prevent endogenous synthesis of adenylic acid. With the aid of this system, a new series of human-mouse hybrids has been prepared between wild type human diploid fibroblasts and an enzyme-deficient mouse line. Survival of the hybrids depended upon the presence of the APRT, which was shown to have the isoelectric pH characteristic of the human enzyme and not that of the mouse. Reduced hybrids containing the enzyme lacked all human biarmed chromosomes, so that unless a rearrangement had occurred, the aprt gene must be located on an acrocentric chromosome. The hybrid cells became APRT- with a frequency of 2 × 10-3, probably by loss of the human aprt chromosome. The APRT- progeny could be obtained selectively by growth in medium containing fluoroadenine.

Full text

PDF
82

Images in this article

84Image
on p.84
84Image
on p.84
84Image
on p.84

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkins J. H., Gartler S. M. Development of a nonselective technique for studying 2, 6, diaminopurine resistance in an established murine cell line. Genetics. 1968 Dec;60(4):781–792. doi: 10.1093/genetics/60.4.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BREITMAN T. R. The feedback inhibition of thymidine kinase. Biochim Biophys Acta. 1963 Jan 8;67:153–155. doi: 10.1016/0006-3002(63)91807-9. [DOI] [PubMed] [Google Scholar]
  3. Coon H. G., Weiss M. C. A quantitative comparison of formation of spontaneous and virus-produced viable hybrids. Proc Natl Acad Sci U S A. 1969 Mar;62(3):852–859. doi: 10.1073/pnas.62.3.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gale G. R., Ostrander W. E., Atkins L. M. Effects of alanosine on purine and pyrimidine synthesis. Biochem Pharmacol. 1968 Sep;17(9):1823–1832. doi: 10.1016/0006-2952(68)90098-1. [DOI] [PubMed] [Google Scholar]
  5. Gale G. R., Schmidt G. B. Mode of action of alanosine. Biochem Pharmacol. 1968 Mar;17(3):363–368. doi: 10.1016/0006-2952(68)90246-3. [DOI] [PubMed] [Google Scholar]
  6. HARRIS M., RUDDLE F. H. Clone strains of pig kidney cells with drug resistance and chromosomal markers. J Natl Cancer Inst. 1961 Jun;26:1405–1411. [PubMed] [Google Scholar]
  7. Kano K., Baranska W., Knowles B. B., Koprowski H., Milgrom F. Surface antigens on interspecies hybrid cells. J Immunol. 1969 Nov;103(5):1050–1060. [PubMed] [Google Scholar]
  8. Kelley W. N., Levy R. I., Rosenbloom F. M., Henderson J. F., Seegmiller J. E. Adenine phosphoribosyltransferase deficiency: a previously undescribed genetic defect in man. J Clin Invest. 1968 Oct;47(10):2281–2289. doi: 10.1172/JCI105913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kelley W. N., Rosenbloom F. M., Henderson J. F., Seegmiller J. E. A specific enzyme defect in gout associated with overproduction of uric acid. Proc Natl Acad Sci U S A. 1967 Jun;57(6):1735–1739. doi: 10.1073/pnas.57.6.1735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kusano T., Wang R., Pollack R., Green H. Human-mouse hybrid cell lines and susceptibility to polio virus. II. Polio sensitivity and the chromosome constitution of the hybrids. J Virol. 1970 Jun;5(6):682–685. doi: 10.1128/jvi.5.6.682-685.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LIEBERMAN I., OVE P. Enzyme studies with mutant mammalian cells. J Biol Chem. 1960 Jun;235:1765–1768. [PubMed] [Google Scholar]
  12. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  13. Littlefield J. W., Goldstein S. Some aspects of somatic cell hybridization. In Vitro. 1970 Jul-Aug;6(1):21–31. doi: 10.1007/BF02616131. [DOI] [PubMed] [Google Scholar]
  14. Matsuya Y., Green H., Basilico C. Properties and uses of human-mouse hybrid cell lines. Nature. 1968 Dec 21;220(5173):1199–1202. doi: 10.1038/2201199a0. [DOI] [PubMed] [Google Scholar]
  15. Matsuya Y., Green H. Somatic cell hybrid between the established human line D98 (presumptive HeLa) and 3T3. Science. 1969 Feb 14;163(3868):697–698. doi: 10.1126/science.163.3868.697. [DOI] [PubMed] [Google Scholar]
  16. Migeon B. R., Miller C. S. Human-mouse somatic cell hybrids with single human chromosome (group E): link with thymidine kinase activity. Science. 1968 Nov 29;162(3857):1005–1006. doi: 10.1126/science.162.3857.1005. [DOI] [PubMed] [Google Scholar]
  17. Murthy Y. K., Thiemann J. E., Coronelli C., Sensi P. Alanosine, a new antiviral and antitumour agent isolated from a Streptomyces. Nature. 1966 Sep 10;211(5054):1198–1199. doi: 10.1038/2111198a0. [DOI] [PubMed] [Google Scholar]
  18. Ruddle F. H., Chapman V. M., Chen T. R., Klebe R. J. Genetic analysis with man-mouse somatic cell hybrids. Linkage between human lactate dehydrogenase A and B and peptidase B. Nature. 1970 Jul 18;227(5255):251–257. doi: 10.1038/227251a0. [DOI] [PubMed] [Google Scholar]
  19. TODARO G. J., GREEN H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol. 1963 May;17:299–313. doi: 10.1083/jcb.17.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vesterberg O., Svensson H. Isoelectric fractionation, analysis, and characterization of ampholytes in natural pH gradients. IV. Further studies on the resolving power in connection with separation of myoglobins. Acta Chem Scand. 1966;20(3):820–834. doi: 10.3891/acta.chem.scand.20-0820. [DOI] [PubMed] [Google Scholar]
  21. Weiss M. C. Further studies on loss of T-antigen from somatic hybrids between mouse cells and SV40-transformed human cells. Proc Natl Acad Sci U S A. 1970 May;66(1):79–86. doi: 10.1073/pnas.66.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weiss M. C., Green H. Human-mouse hybrid cell lines containing partial complements of human chromosomes and functioning human genes. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1104–1111. doi: 10.1073/pnas.58.3.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES