Development of a highly efficient recombinant system for Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV) and findings about baculoviral replication in absence of the essential gene orf1629

HAASE, SANTIAGO; FABRE, MA. LAURA; ROMANOWSKI, VÍCTOR; FERRELLI, MA. LETICIA

Congreso; 49th Annual Meeting of the Society for Invertebrate Pathology International Congress on Invertebrate Pathology and Microbial Control; 2016

Developmentof a highly efficient recombinant system for Anticarsia gemmatalis MultipleNucleopolyhedrovirus (AgMNPV) and findings about baculoviral replication inabsence of the essential gene orf1629.Authors: ML Fabre, S Haase, V RomanowskiInstituto de Biotecnología y Biología Molecular (IBBM) UNLP-CONICET,Argentina.Presentation category: Poster Anticarsiagemmatalis multiple nucleopolyhedronvirus (AgMNPV)is one of the most extensively used viral pesticides, but its performance as abiological control agent in more temperate climates requires the improvement ofits biopesticidal properties. Genetic modification is a powerful strategy toimprove this properties in AgMNPV including heterologous genes in the viralDNA. In our laboratory, we have previously developed a homologous recombination(HR) system to modify AgMNPV genome based on HR in cultured insect cells.Nevertheless, recombinant AgMNPV isolation has proven to be time-consuming. Inorder to simplify this procedure, we developed a novel recombination systembased on a defective AgMNPV (AgMNPV-Δ1629) lacking an essential gene (orf1629)that replicates in a transgenic insect cell line expressing this gene(UFLAg286-1629). By co-transfection of non-transgenic cells (UFLAg286) withparental AgMNPV-Δ1629 DNA and a transfer plasmid (containing orf1629), recombinant baculoviruses arerecovered. HR restoringorf1629 and allows insertion of pesticidalgenes and/or a marker genes under the control of strong viral promoters. Weconstructed AgMNPV-Δ1629 and demonstrated the complementation capacity of thetransgenic cell line. After this, we tested the system by isolating arecombinant virus carrying the green fluorescent protein gene. In addition,novel results were found about the orf1629 gene indicating thatbaculoviral DNA replicates efficiently in absence of this essential gene andthat occlusion bodies appear in transfected cells and confirmed that this geneis essential for budded virus formation.