The synthetic plasmid, pSV2-gpf, was transfected into the cultured fish cells (RBCF-1 line) using polycation and dimethyl sulphoxide. The maximum transfection frequency estimated by colony number in the selection medium was 585 transfectants/5x105 treated cells per 50 ng plasmid DNA. The isolated transfectants expressed the xanthine guanine phosphoribosyltransferase (XGPRT) activity encoded by the plasmid DNA associated with the promoter of simian virus 40 (SV40). The resistance to mycophenolic acid and the XGPRT activity of every transfectant examined were stable, indicating the possibility that pSV2-gpt was integrated into the genomic DNA of the host fish cells. Our results, in addition to those already reported for mammalian and avian cell lines, indicate that the promoter in the early region of SV40 can function in cultured fish cells. This success in obtaining cultured fish cells with a dominant selective marker will provide a useful clue for somatic cell genetic studies of fish in the future.
Transfection and stable expression of a dominant selective marker Ecogpt in a cultured cell line of the fish, Carassius auratus
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K. ISA, A. SHIMA; Transfection and stable expression of a dominant selective marker Ecogpt in a cultured cell line of the fish, Carassius auratus. J Cell Sci 1 September 1987; 88 (2): 219–224. doi: https://doi.org/10.1242/jcs.88.2.219
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