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

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 dip...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 68; no. 1; pp. 82 - 86
Main Authors Kusano, Toshihisa, Long, Cedric, Green, Howard
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences of the United States of America 01.01.1971
National Acad Sciences
Subjects
Acrocentric chromosomes
Adenine - metabolism
Animals
Biological Sciences: Genetics
Cell Line - enzymology
Chromosomes
Culture Techniques
Cultured cells
Cytogenetics
Diploidy
Enzymes
Fibroblasts
Fibroblasts - enzymology
Genes
Genes, Regulator
Human chromosomes
Humans
Hybrid cells
Hybridity
Hybridization, Genetic
Hydrogen-Ion Concentration
Mice
Transferases - biosynthesis
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Summary: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.
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Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, Mass. 02139
Present address: Research Institute for Tuberculosis, Leprosy, and Cancer, Tohoku University, Sendai, Japan.
Postdoctoral Fellow of the National Cancer Institute. Present address: Flow Laboratories, Rockville, Maryland.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.68.1.82