Alternative codon usage of PRRS virus ORF5 gene increases eucaryotic expression of GP5 glycoprotein and improves immune response in challenged pigs
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11 juillet 2017
- handle: 10670/1.grwcnp
- doi: 10.1016/j.vaccine.2005.03.012
Mots-clés
PRRSV Porcine arterivirus GP5 protein Codon bias Synthetic gene Expression Adenovirus vectors ImmunizationSujets proches
Domestic pig Pigs Sus domesticus Pig Sus domestica Pig farming Hogs Sus scrofa domesticus Sus scrofa domesticaCiter ce document
Ali Kheyar et al., « Alternative codon usage of PRRS virus ORF5 gene increases eucaryotic expression of GP5 glycoprotein and improves immune response in challenged pigs », Papyrus : le dépôt institutionnel de l'Université de Montréal, ID : 10.1016/j.vaccine.2005.03.012
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Résumé
Pigs exposed to GP5 protein of PRRSV by means of DNA immunization develop specific neutralizing and protecting antibodies. Herein, we report on the consequences of codon bias, and on the favorable outcome of the systematic replacement of native codons of PRRSV ORF5 gene with codons chosen to reflect more closely the codon preference of highly expressed mammalian genes. Therefore, a synthetic PRRSV ORF5 gene (synORF5) was constructed in which 134 nucleotide substitutions were made in comparison to wild-type gene (wtORF5), such that 59% (119) of wild-type codons were replaced with known preferable codons in mammalian cells. In vitro expression in mammalian cells of synORF5 was considerably increased comparatively to wtORF5, following infection with tetracycline inducible replication-defective human adenoviral vectors (hAdVs). After challenge inoculation, SPF pigs vaccinated twice with recombinant hAdV/synORF5 developed earlier and higher antibody titers, including virus neutralizing antibodies to GP5 than pigs vaccinated with hAdV/wtORF5. Data obtained from animal inoculation studies suggest direct correlation between expression levels of immunogenic structural viral proteins and immune response.