Establishment of a Chinook salmon cell line with an inducible gene expression system
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We have isolated a stable recombinant cell line CHSE-TOF5 derived from the Chinook salmon (Oncorhynchus tshawytscha) embryo cells for use as an inducible expression system. The cells were transfected with the pTet-Off plasmid from the Tet On/Off Clontech system, carrying a G418 resistance gene. Several G418-resistant clones were subcultured and characterised by quantitative PCR (qPCR) and by transient transfection. The level of expression of transcriptional activator was measured by qPCR in a number of isolated clones, and transient transfection with a pTRE2-hyg-LUC plasmid was used to evaluate the inducibility of these clones. A clone was selected for its relative fast cell growth and good level of inducibility. This genetically engineered cell line is a valuable tool for the fish research community especially in research areas investigating the biological function of proteins from fish or fish pathogens.
KeywordsBiomaterials Cell culture Fish cell line Tet-off
- McBeath A. J.; Snow M.; Secombes C. J.; Ellis A. E.; Collet B. Expression kinetics of interferon and interferon-induced genes in Atlantic salmon (Salmo salar) following infection with infectious pancreatic necrosis virus and infectious salmon anaemia virus. Fish Shellfish Immunol. 22: 230–241; 2007.PubMedCrossRefGoogle Scholar