Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus
Here, we developed a diagnostic ELISA for foot-and-mouth disease using recombinant occlusion bodies (rOBs) of baculovirus. We fused Δ3AB1–3, a polypeptide derived from non-structural proteins of foot-and-mouth disease virus, to polyhedrin (POLH), the major constituent of OBs, under polh promoter. To...
| Main Authors: | , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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Springer
2019
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| Online Access: | http://hdl.handle.net/20.500.12123/4459 https://doi.org/10.1007/s13205-018-1482-x |
| _version_ | 1855483436948520960 |
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| author | Diez, Michay Trotta, Myrian Vanesa Alfonso, Victoria Taboga, Oscar Alberto Lopez, Maria Gabriela |
| author_browse | Alfonso, Victoria Diez, Michay Lopez, Maria Gabriela Taboga, Oscar Alberto Trotta, Myrian Vanesa |
| author_facet | Diez, Michay Trotta, Myrian Vanesa Alfonso, Victoria Taboga, Oscar Alberto Lopez, Maria Gabriela |
| author_sort | Diez, Michay |
| collection | INTA Digital |
| description | Here, we developed a diagnostic ELISA for foot-and-mouth disease using recombinant occlusion bodies (rOBs) of baculovirus. We fused Δ3AB1–3, a polypeptide derived from non-structural proteins of foot-and-mouth disease virus, to polyhedrin (POLH), the major constituent of OBs, under polh promoter. To further assess the most convenient strategy to improve yields, we designed two recombinant baculoviruses, vPOLH and vPOLHE44G. These carried the sequence of the fusion protein POLH-Δ3AB1–3 with an additional copy in cis of polh or polhE44G, respectively, under p10 promoter. Our results show that both viruses expressed POLH-Δ3AB1–3, which was detected by western blot in purified rOBs with anti-POLH and anti-3AB1 antibodies. We also found that vPOLHE44G produced larger polyhedra and a significant increase of antigen yield (p < 0.01). Furthermore, the chimeric protein POLH-Δ3AB1–3 was recognized by sera from experimentally infected animals, showing that translational fusion to POLH does not alter the antigenicity of Δ3AB1–3. Finally, the rOBs were successfully used in an ELISA test to differentiate infected from vaccinated animals. Taken together, these results demonstrate the great potential of rOBs to develop diagnostic schemes adaptable to animal infectious diseases. |
| format | Artículo |
| id | INTA4459 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA44592019-02-18T16:32:40Z Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus Diez, Michay Trotta, Myrian Vanesa Alfonso, Victoria Taboga, Oscar Alberto Lopez, Maria Gabriela Baculovirus Virus Fiebre Aftosa Aphthovirus ELISA Occlusion Bodies Foot and Mouth Disease Virus Polyhedrin FMDV DIVA Diagnostic Cuerpos de Oclusión Here, we developed a diagnostic ELISA for foot-and-mouth disease using recombinant occlusion bodies (rOBs) of baculovirus. We fused Δ3AB1–3, a polypeptide derived from non-structural proteins of foot-and-mouth disease virus, to polyhedrin (POLH), the major constituent of OBs, under polh promoter. To further assess the most convenient strategy to improve yields, we designed two recombinant baculoviruses, vPOLH and vPOLHE44G. These carried the sequence of the fusion protein POLH-Δ3AB1–3 with an additional copy in cis of polh or polhE44G, respectively, under p10 promoter. Our results show that both viruses expressed POLH-Δ3AB1–3, which was detected by western blot in purified rOBs with anti-POLH and anti-3AB1 antibodies. We also found that vPOLHE44G produced larger polyhedra and a significant increase of antigen yield (p < 0.01). Furthermore, the chimeric protein POLH-Δ3AB1–3 was recognized by sera from experimentally infected animals, showing that translational fusion to POLH does not alter the antigenicity of Δ3AB1–3. Finally, the rOBs were successfully used in an ELISA test to differentiate infected from vaccinated animals. Taken together, these results demonstrate the great potential of rOBs to develop diagnostic schemes adaptable to animal infectious diseases. Instituto de Biotecnología Fil: Diez, Michay. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Trotta, Myrian Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2019-02-18T16:28:56Z 2019-02-18T16:28:56Z 2018-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/4459 2190-572X 2190-5738 (Online) https://doi.org/10.1007/s13205-018-1482-x eng info:eu-repo/semantics/restrictedAccess application/pdf Springer 3 Biotech 8 : 457. (November 2018) |
| spellingShingle | Baculovirus Virus Fiebre Aftosa Aphthovirus ELISA Occlusion Bodies Foot and Mouth Disease Virus Polyhedrin FMDV DIVA Diagnostic Cuerpos de Oclusión Diez, Michay Trotta, Myrian Vanesa Alfonso, Victoria Taboga, Oscar Alberto Lopez, Maria Gabriela Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus |
| title | Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus |
| title_full | Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus |
| title_fullStr | Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus |
| title_full_unstemmed | Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus |
| title_short | Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus |
| title_sort | recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot and mouth disease virus |
| topic | Baculovirus Virus Fiebre Aftosa Aphthovirus ELISA Occlusion Bodies Foot and Mouth Disease Virus Polyhedrin FMDV DIVA Diagnostic Cuerpos de Oclusión |
| url | http://hdl.handle.net/20.500.12123/4459 https://doi.org/10.1007/s13205-018-1482-x |
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