Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)
- 27 January 2012
- journal article
- Published by Zhejiang University Press in Journal of Zhejiang University-SCIENCE B
- Vol. 13 (2), 111-117
- https://doi.org/10.1631/jzus.b1100131
Abstract
In the late phase of Bombyx mori nucleopolyhedrovirus (BmNPV) infection, a large amount of polyhedra appear in the infected cell nucleolus, these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment. To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV, two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus (polh+) Bac-to-Bac system, designated as vBmBac(polh+)-enhanced green fluorescent protein (EGFP) and vBmBac(polh+)-LacZ, which can express the polyhedrin and foreign protein simultaneously. Light microscopy analysis showed that all viruses produced polyhedra of normal appearance. Green fluorescence can be apparently detected on the surface of the vBmBac(polh+)-EGFP polyhedra, but not the BmNPV polyhedra. Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra. As expected, the vBmBac(polh+)-LacZ polyhedra contained an amount of LacZ and had a higher β-galactosidase activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra. This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins.Keywords
This publication has 17 references indexed in Scilit:
- The atomic structure of baculovirus polyhedra reveals the independent emergence of infectious crystals in DNA and RNA virusesProceedings of the National Academy of Sciences of the United States of America, 2009
- How baculovirus polyhedra fit square pegs into round holes to robustly package virusesThe EMBO Journal, 2009
- Structure-based targeting of bioactive proteins into cypovirus polyhedra and application to immobilized cytokines for mammalian cell cultureBiomaterials, 2009
- Characterization of a virion occlusion-defective Autographa californica multiple nucleopolyhedrovirus mutant lacking the p26, p10 and p74 genesJournal of General Virology, 2009
- Efficient production of canine interferon-alpha in silkworm Bombyx mori by use of a BmNPV/Bac-to-Bac expression systemApplied Microbiology and Biotechnology, 2008
- The molecular organization of cypovirus polyhedraNature, 2007
- Immobilization of diverse foreign proteins in viral polyhedra and potential application for protein microarraysProteomics, 2006
- The Baculoviruses Occlusion‐Derived Virus: Virion Structure and FunctionPublished by Elsevier BV ,2006
- Chromophore Formation in Green Fluorescent ProteinBiochemistry, 1997
- An Enhanced Green Fluorescent Protein Allows Sensitive Detection of Gene Transfer in Mammalian CellsBiochemical and Biophysical Research Communications, 1996