Infección oral de larvas de lepidópteros con baculovirus para la expresión de peroxidasa recombinante.

ROMERO L.; TARGOVNIK A.; LEVIN G.; CASCONE O.; MIRANDA MV

CABA Argentina

Congreso; IX Congreso Argentino de Virología; 2008

 Two different strategies to generate recombinant baculovirus suitable for oral infection for horseradish peroxidase (HRP) production were assessed: 1) Production of mixed polyhedra by co-infection with wild type AcMNPV and AcMNPV HRP+/occ-. 2) AcMNPV HRP+/occ+ construction by homologous recombination between transfer vector pAcGP67-HRP and bAcGOZA to produce occluded baculovirus. Rachiplusia nu and Spodoptera frugiperda larvae were fed with an artificial diet contaminated with polyhedra (1x106 pol/100 mg diet). Multiplicities of infection wild:recombinant virus 2:10 and 2:20 were the best conditions for R. nu infection with mixed polyhedra, where HRP activity in haemolymph at day 4 post-infection was 78.5 ± 10.3 U ml-1 (31.1 ± 4.5 mg l-1) and 40.3 ± 20.2 U ml-1 (16.2 ± 8.4 mg l-1) respectively. However, the enzyme activity obtained with AcMNPV HRP+/occ+ was significantly higher: 120.5 ± 15.5 U ml-1 (47.5 ± 6.5 mg l-1), (P < 0.05). S. frugiperda larvae evidenced lower susceptibility to oral infection in comparison with R. nu. According to these results, R. nu larva is the best choice for HRP production by oral infection with AcMNPV HRP+/occ+. This strategy is more effective and practical than HRP production via mixed polyhedra.