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Stable Protein Expression in Mammalian Cells Using Baculoviruses

  • Andreas Lackner
  • Emanuel Kreidl
  • Barbara Peter-Vörösmarty
  • Sabine Spiegl-Kreinecker
  • Walter Berger
  • Michael GruschEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 801)

Abstract

The baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) has been widely used in biotechnology for protein expression in insect cells. Baculoviruses use arthropods as their natural hosts and are unable to replicate in mammalian cells. However, AcMNPV is able to enter many mammalian cell types and can be used for transgene expression if engineered to contain suitable expression cassettes. In this chapter, we describe the construction and application of a recombinant baculovirus containing a bicistronic expression cassette that can be used for stable protein expression in mammalian cells. As an example, the generation of glioblastoma and hepatocellular carcinoma cell lines stably expressing green fluorescent protein after puromycin selection is shown.

Key words

Baculovirus BacMam Stable protein expression IRES Green fluorescent protein 

Notes

Acknowledgments

We thank Katharina Leopold for help with the plasmids and Irene Herbacek for performing the flow cytometry analyses.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Andreas Lackner
    • 1
  • Emanuel Kreidl
    • 1
  • Barbara Peter-Vörösmarty
    • 1
  • Sabine Spiegl-Kreinecker
    • 2
  • Walter Berger
    • 1
  • Michael Grusch
    • 1
    Email author
  1. 1.Department of Medicine I, Institute of Cancer ResearchMedical University of ViennaViennaAustria
  2. 2.Department of NeurosurgeryWagner Jauregg HospitalLinzAustria

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