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Title: DIFFERENTIALLY EXPRESSED MIDGUT TRANSCRIPTS IN CULICOIDES SONORENSIS (DIPTERA: CERATOPOGONIDAE) FOLLOWING ORBIVIRUS (REOVIRIDAE) ORAL FEEDING

Author
item Campbell, Corey
item Wilson, William

Submitted to: Insect Molecular Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/19/2002
Publication Date: 12/1/2002
Citation: Campbell, C.L., Wilson, W.C. 2002. Differentially expressed midgut transcripts in culicoides sonorensis (diptera: ceratopogonidae) following orbivirus (reoviridae) oral feeding. Insect Molecular Biology. 11:595-604.

Interpretive Summary: Understanding insect's gene expression response to a virus infection may aid design of control measures for many diseases. Two different approaches were used to identify biting midge genes that are more highly expressed 1 day following a feeding of epizootic hemorrhagic disease virus. Quantitative nucleic acid amplification methods confirmed seven genes out of over 400 candidates to be more abundant in infected midguts than controls. Virus entry into midgut cells was confirmed by identification of a gene induced by double-stranded RNA that is consistent with the virus genomic RNA. Other genes identified in this assay coded for receptors involved in cell differentiation, such as CsLAR and CsFZ2. Other genes were identified which may control protein translation, such as CseIF3, CseIF5a, and CsRPS6. Quantitative amplification methods were also used to compare variations in gene expression levels 1- 3 days after feeding.

Technical Abstract: Understanding the vector insect's gene expression response to a virus infection may aid design of control measures for arbovirus diseases. Differential display and a subtractive library were used to identify Culicoides sonorensis midgut transcripts more abundant 1 day following per os introduction of epizootic hemorrhagic disease virus. Of 400+ candidates screened, real-time PCR confirmed seven transcripts more abundant in infected midguts than controls. Viral entry into midgut cells was confirmed by isolation of RNA editase CsRED1, induced by dsRNA. Transcripts coding for receptors involved in cell differentiation included CsLAR, a putative protein tyrosine phosphatase, and CsFZ2, homologous to the wingless receptor in D. melanogaster. Transcripts encoding putative translation machinery components included CseIF3, CseIF5a, and CsRPS6. Relative transcript abundance was assayed 1- 3 days post-feeding.