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The Regulation of Innate Immune Responses to Endoparasites in Lepidopteran and Dipteran Larvae
B.A. Webb, S. Govind
Department of Entomology
Project Description
Webb Lab: The functional analysis of the CsIV vankyrin genes is proceeding. By quantitative RT-PCR we have documented that the viral vankyrin genes regulate some but not all anti-microbial peptide genes by gene specific quantitative PCR. We have also developed a H. virescens microarray in collaboration with Dr. Kent Shelby (USDA:Missouri Biocontrol Lab). This array has approximately 15,000 unigenes and is the first such array developed for H. virescens to our knowledge. Teh development of this array was not anticipated in our proposal but will enable us to perform the analyses of H.v. immune genes in much more depth than initially proposed. It will also be of use to other researchers working with this insect. We performed initial hybridizations with immune challenged cells which we are currently evaluating. Finally, we have shown that the CsIV vankyrin genes interact directly with the Nf-kb signalling pathway using mammalian based reporter assays.
Govind Lab: Using real-time PCR and genetics, we have confirmed that parasitoid-induced activation of antimicrobial gene expression in Drosophila is dependent on Toll-NF-κB signaling. We are cloning CsIV pVank1 and pVank3 in the Drosophila transformation vector to express the respective proteins in transgenic flies.
Impact
The first significant outcome of this project is the development of a microarray for H. virescens. This opportunity resulted from the PI's efforts to identify H. virescens immune genes that had been sequenced but were not in public databases. Ultimately this line of inquiry resulted in a donation of approximately 40,000 EST sequences and a similar number of aphid EST sequences to the University of Kentucky by a FMC corporation. These sequences were made available and prompted other investigators to donate sequences to the community. The sequences were deposited to public databases and are in use by researchers.
Publications
Schlenke, T.A., Morales, J., Govind, S., Clark, A.G. (2007) Contrasting infection strategies in generalist and specialist wasp parasitoids of Drosophila melanogaster. PLoS Pathogens. PLoS Pathog 3(10): e158.
Govind, S. (2008) Innate immunity in Drosophila: Pathogens and pathways. Insect Science 15:29-43.