Quantitative measurement of mammalian chromosome mitotic loss rates using the green fluorescent protein.
We have measured the mitotic loss rates of mammalian chromosomes in cultured cells. The green fluorescent protein (GFP) gene was incorporated into a non-essential chromosome so that cells containing the chromosome fluoresced green, while those lacking it did not. The proportions of fluorescent and non-fluorescent cells were measured by fluorescence activated cell sorter (FACS) analysis. Loss rates ranged from 0.005% to 0.20% per cell division in mouse LA-9 cells, and from 0.02% to 0.40% in human HeLa cells. The rate of loss was elevated by treatment with aneugens, demonstrating that the system rapidly identifies agents which induce chromosome loss in mammalian cells.
CRC Chromosome Molecular Biology Group, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
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EM
Burns
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Christopoulou
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P
Corish
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Tyler-Smith
D000782:Aneuploidy; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D002021:Buffers; D002110:Caffeine / Q000494:pharmacology; D002459:Cell Fusion; D002869:Chromosome Aberrations; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D003703:Demecolcine / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D005947:Glucose / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D008322:Mammals; D051379:Mice; D008938:Mitosis / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D015739:Nocodazole / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D015003:Yeasts
D000970:Antineoplastic Agents; D000972:Antineoplastic Agents, Phytogenic; D002021:Buffers; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D010726:Phosphodiesterase Inhibitors; D049452:Green Fluorescent Proteins; D002110:Caffeine; D005047:Etoposide; D005947:Glucose; D015739:Nocodazole; D003703:Demecolcine