MERZOUKI, Abderrazzak JEAN, Myriam HENRICHON, Marc GUERTIN, Claude
Title
Production of Anti-EGT Polyclonal Antibodies Using The Genetic Immunisation Method.
Abstract Text
The granulovirus of Choristoneura fumiferana (ChfuGV) is a natural pathogen of the spruce budworm larvae, the principal pest of coniferous forests. The molecular and immunological characterization of viral proteins leads to a better understanding of virus/host interactions. The ecdysteoid-glucosyl-transferase (EGT) protein is an enzyme coded by the viral gene egt. The EGT enzyme disrupts the hormonal balance of the insect host’s molting hormones, preventing infected larvae from molting or pupating. In this project the EGT protein is analyzed for its molecular and immunological properties. The egt gene was amplified by PCR and cloned to the pCR2.1 vector, then subcloned to the pcDNA3 eucaryotic expression vector using its BamHI and XbaI restriction sites. The obtained pcDNA3/EGT vector was amplified in E. coli XL1-Blue cells, purified and used for the genetic immunization of Balb/c mice so as to raise anti-EGT polyclonal antibodies. The level of the production of these antibodies will be evaluated relatively to the use of the murine interleukin-2 (IL-2), produced by co-injected pcDNA/IL2 vector. The anti-EGT antibodies raised this way will be used in immunological screening of infected larvae histological cuts for the presence of the EGT protein. The results of this study should help to better understand the role of the EGT protein in the pathogenicity of the ChfuGV virus.
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