A promising genomic transfectant into Xeroderma pigmentosum group A with highly amplified mouse DNA and intermediate UV resistance turns revertant.
Following transfection of genomic mouse DNA into an SV40 transformed fibroblast cell line from a patient with Xeroderma pigmentosum (complementation group A, XPA), a single UV resistant cell clone was isolated out of a total of 10(4) independent transfectants. The recipient XPA cell line has as yet not produced spontaneous revertants among 2.2 x 10(8) cells. The isolated cell clone contains 50-70 kb of mouse sequences which are heavily amplified (500-fold), and has acquired both intermediate resistance to UV killing and intermediate unscheduled DNA synthesis (UDS) capacity. By continued passage without selective pressure, cells were generated, which had lost both the dominant marker gene and repetitive mouse sequences. Single colonies of these cells were still intermediately resistant to UV suggesting that either undetected unique mouse DNA had segregated from the bulk of repetitive DNA, or, more likely, that the initially isolated transfectant was a spontaneous revertant. This documents that a persuasive clone isolated can still be a false positive (spontaneous revertant) and that an extremely laborious approach may lead into a dead end.
Blum
M
M
Kernforschungszentrum Karlsruhe, Institut für Genetik und Toxikologie, FRG.
Baumann
I
I
Lohrer
H
H
Rahmsdorf
H J
HJ
Herrlich
P
P
D000818:Animals; D001483:Base Sequence / Q000528:radiation effects; D015139:Blotting, Southern; D002461:Cell Line, Transformed; D004247:DNA / Q000528:radiation effects*; D004252:DNA Mutational Analysis; D005784:Gene Amplification; D006801:Humans; D051379:Mice; D010641:Phenotype; D014162:Transfection / Q000528:radiation effects*; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D004247:DNA