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Amplicon structure in multidrug-resistant murine cells: a nonrearranged region of genomic DNA corresponding to large circular DNA.

Artikel i vetenskaplig tidskrift
Författare F Ståhl
Yvonne Wettergren
G Levan
Publicerad i Molecular and cellular biology
Volym 12
Nummer/häfte 3
Sidor 1179-87
ISSN 0270-7306
Publiceringsår 1992
Publicerad vid Institutionen för anatomi och cellbiologi
Sidor 1179-87
Språk en
Länkar www.ncbi.nlm.nih.gov/entrez/query.f...
Ämnesord Animals, Blotting, Southern, DNA, genetics, DNA, Circular, genetics, Drug Resistance, genetics, Electrophoresis, Gel, Pulsed-Field, Gene Amplification, Mice, Restriction Mapping, Tumor Cells, Cultured
Ämneskategorier Cellbiologi

Sammanfattning

Multidrug resistance (MDR) in tumor cell lines is frequently correlated with amplification of one or more mdr genes. Usually the amplified domain also includes several neighboring genes. Using pulsed-field gel electrophoresis, we have established a restriction map covering approximately 2,200 kb in the drug-sensitive mouse tumor cell line TC13K. The mapped region is located on mouse chromosome 5 and includes the three mdr genes, the gene for the calcium-binding sorcin protein, and a gene with unknown function designated class 5. Long-range maps of the amplified DNA sequences in five of six MDR sublines that had been independently derived from TC13K generally displayed the same pattern as did the parental cell line. All six MDR sublines exhibited numerous double minutes, and one of them displayed a homogeneously staining region in a subpopulation. Large circular molecules, most likely identical to one chromatid of the double minutes, were detected in four of the sublines by linearization with gamma irradiation. The size of the circles was about 2,500 kb, which correlated to a single unit of the amplified domain. We therefore propose that in four independent instances of MDR development, a single unit of about 2,500 kb has been amplified in the form of circular DNA molecules. The restriction enzyme map of the amplified unit is unchanged compared with that of the parental cell line, whereas the joining sites of the circular DNA molecules are not identical but are in the same region.

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