High-level expression of recombinant 3AB1 non-structural protein from FMDV in insect larvae.
For its potential usefulness in diagnosis, the non-structural protein 3AB1 from foot-and-mouth disease virus was expressed as a soluble protein by using Autographa californica nuclear polyhedrosis virus as a vector. The 3AB1 coding sequence was introduced into AcNPV genome via pBAcPAK3AB1 transfer vector to originate Ac3AB1 recombinant baculovirus of phenotype occ-. Rachiplusia nu larvae were injected with supernatants of Sf9 cells infected with Ac3AB1 and 5 days post-infection total protein extracts were obtained. An intense band of approximately 21.5 kDa was observed when total larvae extracts were SDS-PAGE resolved and the recombinant protein detected by an FMDV-infected guinea pig serum. ELISA tests and Western blot experiments were carried out using sera both from FMDV-infected cattle and from vaccinated animals. The recombinant protein was only recognized by sera from infected animals, suggesting that this method of production in insect larvae could be applied to an efficient mass production of proteins of diagnostic interest.
López
María Gabriela
MG
Instituto de Biotecnología, CICVyA, INTA, Castelar, CC25 (1712), Buenos Aires, Argentina.
Peralta
Andrea
A
Berinstein
Analía
A
Fondevila
Norberto
N
Carrillo
Elisa
E
Taboga
Oscar
O
D000818:Animals; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D007814:Larva; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018411:Spodoptera; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
C109754:3ABC protein, virus; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins