期刊
JOURNAL OF VIROLOGICAL METHODS
卷 187, 期 2, 页码 406-412出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2012.11.011
关键词
Foot-and-mouth disease virus; Recombinant baculovirus; Empty capsids; Protein processing; Frameshift; 3C protease; Vaccine
资金
- Department for Environment, Food and Rural Affairs (DEFRA) [2811]
- BBSRC [BB/J004561/1]
- John Innes Foundation
- Biotechnology and Biological Sciences Research Council [BBS/E/J/000CA326, BBS/E/I/00001716, BBS/B/06547, BBS/E/I/00001703, BBS/E/I/00001494] Funding Source: researchfish
- Medical Research Council [G1100525, G1000769, G1000099] Funding Source: researchfish
- BBSRC [BBS/E/I/00001703, BBS/E/I/00001716, BBS/E/I/00001494, BBS/E/J/000CA326] Funding Source: UKRI
- MRC [G1000769, G1000099] Funding Source: UKRI
Foot-and-mouth disease virus (FMDV) is a significant economically and distributed globally pathogen of Artiodactyla. Current vaccines are chemically inactivated whole virus particles that require large-scale virus growth in strict bio-containment with the associated risks of accidental release or incomplete inactivation. Non-infectious empty capsids are structural mimics of authentic particles with no associated risk and constitute an alternate vaccine candidate. Capsids self-assemble from the processed virus structural proteins, VP0, VP3 and VP1, which are released from the structural protein precursor P1-2A by the action of the virus-encoded 3C protease. To date recombinant empty capsid assembly has been limited by poor expression levels, restricting the development of empty capsids as a viable vaccine. Here expression of the FMDV structural protein precursor P1-2A in insect cells is shown to be efficient but linkage of the cognate 3C protease to the C-terminus reduces expression significantly. Inactivation of the 3C enzyme in a P1-2A-3C cassette allows expression and intermediate levels of 3C activity resulted in efficient processing of the P1-2A precursor into the structural proteins which assembled into empty capsids. Expression was independent of the insect host cell background and leads to capsids that are recognised as authentic by a range of anti-FMDV bovine sera suggesting their feasibility as an alternate vaccine. (C) 2012 Elsevier B.V. All rights reserved.
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