Abstract
The firefly luciferase gene was introduced into the Semliki Forest virus (SFV) vector and high titer recombinant SFV particles generated. The broad host range of SFV allowed efficient infection and high level expression of four mammalian cell lines growing in serum-free suspension cultures. The incubation temperature had dramatic effects on the level and duration of recombinant protein expression. For example, the luciferase activity was significantly higher in the rodent BHK and CHO cell lines incubated at 33 °C compared to 37 °C when harvested 19 h post-infection. At 33 °C the specific expression levels increased 10-20 fold during prolongation of the post-infection time up to 50 h. In contrast, a significant decrease in luciferase activity was observed from 26 h post-infection for cell cultures incubated at 37 °C. Only a slight temperature effect on luciferase expression was seen in the human cell line HEK293 and no effect was observed for the subclone293(EBNA).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Berglund P, Sjöberg M, Garoff H, Atkins GJ, Sheahan BJ and Liljeström P (1993) Bio/Technology 11: 916–920.
Blasey HD, Aubry J-P, Mazzei G and Bernard AR (1996) Cytotech-nology 18: 183–192.
Blasey HD, Lundstrom K, Tate S and Bernard AR (1996) Cytotech-nology 24: 65–72.
Fong TM, Anderson SA, Yu H, Huang RC and Strader C (1992) Mol Pharmacol 41: 24–30.
Kretzmer G, Rössler B, Hüners M, Lübben H and Reich T (1997) In: Animal Cell Technology. Carrondo MJT (ed.) Kluwer Academic Press The Netherlands, pp. 319–323.
Legendre JY, Trzeciak A, Bohrmann B, Deuschle U, Kitas EA and Supersaxo A (1996) Bioconjugate Chem 8: 57–63.
Lundstrom K, Mills A, Buell G, Allet E, Adami N and Liljestrom P (1994) Eur J Biochem 224: 917–921.
Lundstrom K, Pralong W and Martinou J-C (1997) Apoptosis 2: 189–191.
Mastrangelo AJ and Betenbaugh MJ (1998) Tibtech 16: 88–95.
Nakajima Y, Tsuchida K, Negishi M, Ito S and Naganishi S (1992) J Biol Chem 267: 2437–2442.
Schlaeger E-J (1996) J Immunol Meth 194: 191–199.
Schumpp B and Schlaeger E-J (1990) J Cell Sci 97: 639–647.
Weidemann R, Ludwig A and Kretzmer G (1994) Cytotechnology 15: 111–116.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Schlaeger, EJ., Lundstrom, K. (1998). Effect of temperature on recombinant protein expression in Semliki Forest virus infected mammalian cell lines growing in serum-free suspension cultures. In: Betenbaugh, M.J., Chalmers, J.J., Arathoon, R., Chaplen, F.W.R., Mastrangelo, A.J. (eds) Cell Culture Engineering VI. Current Applications of Cell Culture Engineering, vol 3. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-4786-6_21
Download citation
DOI: https://doi.org/10.1007/978-94-011-4786-6_21
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-6011-0
Online ISBN: 978-94-011-4786-6
eBook Packages: Springer Book Archive