Polydnaviruses have an obligate mutualistic association with some members of the Ichneumonidae and Braconidae families of parasitic wasps. CsIV the prototype Ichnovirus is characterized by a large multisegmented genome which contains four putative gene families that encode multiple gene variants. Functional characterization of the gene families has emphasized using whole organism approaches while tissue culture approaches have largely been neglected. Tissue culture work to date has emphasized expression of a particular CsIV gene or protein in an insect cell line while analysis of whole virus infection has not been done. Preliminary results suggest that CsIV is able to infect and induce apoptosis in Sf9 cells. We believe that tissue culture assays will be invaluble in functionally characterizing members of the CsIV gene families therefore we are developing a tissue culture assay in order to infect insect cells with CsIV and functionally characterize two members of the cys-motif gene family, VHv1.4 and VHv1.1. Recent work by Kim et al. suggests that in-vitro transcription of VHv1.4 a member of the cys-motif gene family has HTIF activity in HI-5 cells. We believe that anaylysis of these two genes in respect to whole virus infection will enable us to obtain a better understanding of their roles during virus infection. Furthermore, development of a tissue culture assay will enable us to perform RNAi assays using three recently completed Litmus 28i constructs that can be used to make dsRNA specific for VHv1.4 and VHv1.1.
Species 1: Lepidoptera Noctuidae Heliothis virescens (tobacco budworm)
Species 2: Hymenoptera Ichneumonidae Campoletis sonorensis
Keywords: polydnavirus, RNA interference
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