Transfection of the dominant selective marker, pSV2neo, into a fish cell line epithelioma papulosum cyprini (EPC).
1991
Araki K. | Nagoya H. | Onozato H.
The synthetic plasmid, pSV2neo, was transfected into a fish epithelium cell line (EPC) by calcium phosphate, electroporation, protoplast fusion and DEAE-dextran methods, which are used for mammalian cultured cells, to determine the optimum transfection method applicable for cultured fish cells. The efficiency of transfection was estimated by counting the number of colonies of stable transformants in a selection medium. 42, 26, and 21 transformants/5x10(6) cells per 30 mug plasmid DNA were obtained by the calcium-phosphate, electroporation, and protoplast fusion methods, respectively. Southern hybridization indicated that the isolated stable transformants had a complete neo gene associated with the promoter and enhancer of SV40. These results suggest that pSV2neo was successfully integrated into the genome of the host fish cells by these methods and that the early promoter and enhancer region of SV40 can function in cultured fish epithelium cells.
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