Advertisement

Establishment of a Chinook salmon cell line with an inducible gene expression system

  • Bertrand Collet
  • Katherine Lester
Report

Abstract

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.

Keywords

Biomaterials Cell culture Fish cell line Tet-off 

References

  1. Collet B.; Lester K. Isolation of an inducible expression cell line in Atlantic salmon Salmo salar. J. Biotech. 154: 209–211; 2011.CrossRefGoogle Scholar
  2. Lannan C. N.; Winton J. R.; Fryer J. L. Fish cell lines: establishment and characterization of nine cell lines from salmonids. In Vitro 20: 671–676; 1984.PubMedCrossRefGoogle Scholar
  3. Martin S. A. M.; Collet B.; MacKenzie S.; Evensen O.; Secombes C. J. Genomic tools for examining immune gene function in salmonid fish. Rev. Fish. Sci. 16: 110–116; 2008.CrossRefGoogle Scholar
  4. 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
  5. Munoz I.; Carrillo M.; Zanuy S.; Gomez A. Regulation of exogenous gene expression in fish cells: an evaluation of different versions of the tetracycline-regulated system. Gene 363: 173–182; 2005.PubMedCrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2011

Authors and Affiliations

  1. 1.Immunology and Infection, Marine Scotland, Marine LaboratoryAberdeenUK

Personalised recommendations