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Repetitive DNA sequences in methotrexate- and methasquin-sensitive and -resistant Chinese hamster cell lines

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Abstract

DNA purified from a Chinese hamster cell line of lung fibroblast origin (DC83F) was analyzed by density gradient centrifugation and by gel electrophoresis after restriction endonuclease digestion in order to fractionate discrete repetitive fractions within the total DNA. No obvious satellite DNAs were resolved using the CsCl or Ag-Cs2SO4 density gradient conditions described herein. However, analysis of the digestion products of a battery of restriction endonucleases indicated that three of these enzymes, EcoR 1, HaeIII, and XhoI, yielded discrete fragments which could be visualized with EtBr staining or identified by scintillation counting of [3H] DNA. DNAs from several highly (≥ hundredfold increased resistance) antifolate-resistant sublines of DC-3F, characterized by a large homogeneously staining region (HSR) in the chromosome complement, were examined with both techniques and compared to the parental, antifolate-sensitive cell line DNA. The density gradient profiles and electrophoretic patterns of restriction endonuclease digests were identical among all the cell lines examined and were indistinguishable from those of the parental DC-3F DNA.

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Author notes

  1. Debra J. Wolgemuth

    Present address: Department of Molecular Cell Biology, The Rockefeller University, 10021, New York, New York

Authors and Affiliations

  1. Laboratories of RNA Synthesis and Regulation and Cellular and Biochemical Genetics, Sloan-Kettering Institute for Cancer Research, 10580, Rye, New York

    Debra J. Wolgemuth, June L. Biedler & Peter W. Melera

Authors
  1. Debra J. Wolgemuth
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  2. June L. Biedler
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  3. Peter W. Melera
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Additional information

This work was supported in part by grants to the Sloan-Kettering Institute and to J.L.B. and P.W.M. by the National Institutes of Health and the Fairchild New Frontiers Fund. Portions of this study were presented at the 18th Annual Meeting of the American Society for Cell Biology held in San Antonio, Texas, November 4–8, 1978.

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Wolgemuth, D.J., Biedler, J.L. & Melera, P.W. Repetitive DNA sequences in methotrexate- and methasquin-sensitive and -resistant Chinese hamster cell lines. Biochem Genet 18, 655–667 (1980). https://doi.org/10.1007/BF00484583

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  • DOI: https://doi.org/10.1007/BF00484583

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