The repair of UV-irradiated plasmids transfected into cultured fish cells.
UV-irradiated plasmids (pNPV B and pBSCATSV) were transfected into RBCF-1 cells derived from a goldfish (Carassius auratus) and into a xeroderma pigmentosum (group A) cell line, XP20SSV. The frequency of stable neor transformation by pNPV B decreased in a dose-dependent manner. However, in spite of large differences in UV sensitivity detected in the colony formation assay, the dose-response curves of RBCF-1 cells and XP20SSV cells were almost the same. The photorecovery (PR) of transforming activity of UV-irradiated plasmids was confirmed in RBCF-1 cells but its extent was much smaller than that observed in the survival assay. The expression of the transfected cat (chloramphenicol acetyltransferase; CAT) gene after 24-h incubation in the dark was much more sensitive to UV irradiation when compared with the stable transformation assay. The extent of PR of cat gene expression in RBCF-1 cells was high and comparable with that of the survival assay. The CAT value of RBCF-1 cells transfected with UV-irradiated plasmids relative to that of unirradiated controls increased as incubation time in the dark after transfection became longer. This suggests that the UV lesions on the plasmids transfected in the RBCF-1 cells were repaired in the dark. The cat gene expression of UV-irradiated plasmids in XP20SSV was very low and independent of incubation time after transfection.
Mitani
H
H
Zoological Institute, Faculty of Science, University of Tokyo, Japan.
Komura
J
J
Shima
A
A
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D006054:Goldfish; D006801:Humans; D007700:Kinetics; D010957:Plasmids / Q000528:radiation effects*; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum
D015500:Chloramphenicol O-Acetyltransferase