Characterization of DNA rearrangements of N-myc gene amplification in three neuroblastoma cell lines by pulsed-field gel electrophoresis.
We characterized N-myc gene amplification in three human neuroblastoma cell lines (IMR-32, TGW, GOTO). Rearrangements in long-range regions surrounding amplified N-myc genes were examined by pulsed-field gel electrophoresis. Since rare-cutting enzymes completely digested DNA at the middle of the N-myc gene, we were able to construct a physical map upstream and downstream of the germline N-myc gene, and to obtain information on restriction sites surrounding amplified N-myc genes. This method enables us to envisage the organization of amplified units over a long range. Digestion patterns differed considerably among the germline and the three cell lines, but were simple in each case. We estimated that the minimal distance between neighboring N-myc genes is at least several hundred kilobases. Our data suggest that amplification units contain several DNA fragments derived from different loci, but that they are homogeneous.
Kato
H
H
Institute for Comprehensive Medical Science, Fujita-Gakuen Health University, Aichi, Japan.
Okamura
K
K
Kurosawa
Y
Y
Kishikawa
T
T
Hashimoto
K
K
D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics*; D004586:Electrophoresis / Q000379:methods; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000235:genetics*; D009857:Oncogenes; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D004247:DNA