Abstract
Although the baculovirus-mediated foreign protein expression system has been extensively documented, system investigation of the optimal infection conditions remains elusive. In this study, a recombinant baculovirus vBmBacJS13-egfp was applied for systematic infection of Bombyx mori nucleopolyhedrovirus (BmNPV). We revealed that incubation time and multiplicity of infection (MOI) were the crucial factors affecting the efficiency of virus infection and protein expression. Given the appropriate MOI, BmNPV infection efficiency was increased with elongation of incubation time; the optimal incubation time for BmNPV entry into host cells was about 4 h. Alternatively, an increasing MOI could shorten the incubation time of virus invasion to achieve similar infection efficiency and heterogenous proteins expression levels. This study laid a foundation for the study of the functional genome of BmNPV and provided important parameters for significant improvement on the expression level of heterogenous proteins using the BmNPV-cell expression system.
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Abbreviations
- BmNPV:
-
Bombyx mori nucleopolyhedrovirus
- AcMNPV:
-
Autographa californica nucleopolyhedrovirus
- BEVS:
-
Baculovirus expression vector system
- ODV:
-
Occlusion-derived virus
- BV:
-
Budding virus
- MOI:
-
Multiplicity of infection
- qPCR:
-
Quantitative real-time PCR
- eGFP:
-
Enhanced green fluorescent protein
- PBS:
-
Phosphate buffer saline
- hpi.:
-
Hours post-infection
- TBS:
-
Tris-buffered saline
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Acknowledgements
We thank Manli Wang at Wuhan Institute of Virology, CAS, for providing the VP39 antibody. We thank Charles Amanze for proofreading this manuscript.
Funding
This work was supported by the National Science Foundation of China (31670152) and Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX20_3114).
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Xu, Y., Liu, N., Yang, F. et al. Optimization of infection parameters improves Bombyx mori nucleopolyhedrovirus invasion efficiency. Biologia 77, 261–267 (2022). https://doi.org/10.1007/s11756-021-00931-6
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DOI: https://doi.org/10.1007/s11756-021-00931-6