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Expression of two heterologous proteins depends on the mode of expression: comparison of in vivo and in vitro methods

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Abstract

The yield of two proteins, avidin and green fluorescent protein (GFP), expressed from a modified Autographa californica nucleopolyhedrovirus (AcMNPV), was compared in Sf9 cell culture monolayer, Sf21 cell suspension culture and intact Spodoptera litura larvae. GFP expressed from the p10 promoter yielded up to 1.5% of total soluble protein in larvae, 20-fold higher than that in monolayer suspension culture. Avidin, expressed from the polh promoter, yielded up to 2.3% of total soluble protein in larvae, 10-fold higher than that in suspension culture and 40-fold higher than that in monolayers. Avidin expression did not affect amounts of GFP in dual-expressing baculovirus compared with those detected from a GFP-only expressing AcMNPV. A biotin-binding assay showed that all avidin expressed in larvae was fully active. Glycosylation patterns of chicken-avidin and Spodoptera-avidin were very similar, though the latter showed a proportion of partially glycosylated material.

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Abbreviations

AcMNPV:

Autographa californica nucleopolyhedrovirus

BEVS:

Baculovirus-based expression vector system

GFP:

Green fluorescent protein

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Acknowledgments

This research was supported by the New Zealand Government Public Good Science Fund (FfRST Contract CO6X0207).

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Authors and Affiliations

  1. Horticulture and Food Research Institute of NZ, Tennant Drive, Palmerston North, 4474, New Zealand

    Laurence N. Gatehouse & John T. Christeller

  2. Horticulture and Food Research Institute of NZ, 120 Mt Albert Rd, Auckland, New Zealand

    Ngaire P. Markwick & Joanne Poulton

  3. Department of Microbiology and Immunology, School of Medical Sciences, University of Otago, P.O. Box 56, Dunedin, 9054, New Zealand

    Vivienne L. Young & Vernon K. Ward

  4. HortResearch, Private Bag 11030, Palmerston North, 4442, New Zealand

    Laurence N. Gatehouse

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  1. Laurence N. Gatehouse
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  2. Ngaire P. Markwick
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  5. Vernon K. Ward
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Correspondence to Laurence N. Gatehouse.

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Gatehouse, L.N., Markwick, N.P., Poulton, J. et al. Expression of two heterologous proteins depends on the mode of expression: comparison of in vivo and in vitro methods. Bioprocess Biosyst Eng 31, 469–475 (2008). https://doi.org/10.1007/s00449-007-0184-2

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  • DOI: https://doi.org/10.1007/s00449-007-0184-2

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