Biochemical Genetics

, Volume 18, Issue 7–8, pp 655–667 | Cite as

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

  • Debra J. Wolgemuth
  • June L. Biedler
  • Peter W. Melera
Article

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, EcoR1, HaeIII, and XhoI, yielded discrete fragments which could be visualized with EtBr staining or identified by scintillation counting of [ 3 H] 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.

Key words

methotrexate resistance restriction endonuclease analysis repetitive DNA HSRs 

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Copyright information

© Plenum Publishing Corporation 1980

Authors and Affiliations

  • Debra J. Wolgemuth
    • 1
  • June L. Biedler
    • 1
  • Peter W. Melera
    • 1
  1. 1.Laboratories of RNA Synthesis and Regulation and Cellular and Biochemical GeneticsSloan-Kettering Institute for Cancer ResearchRye

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