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

✍ Scribed by Debra J. Wolgemuth; June L. Biedler; Peter W. Melera

Publisher
Springer
Year
1980
Tongue
English
Weight
710 KB
Volume
18
Category
Article
ISSN
0006-2928

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✦ Synopsis

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-Cs2SO 4 density gradient conditions described herein. However, analysis of the digestion products of a battery of restriction endonucleases indicated that three of these enzymes, E c o R 1, H a e I I I , and X h o I , 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 D C-3F DNA.

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