We are interested in expressing genes targeted to pathogens in appropriate tissues in mosquitoes. We contend that confining expression of these genes to these tissues should maximize the effects of the anti-pathogenic gene without compromising the fitness of the insect vector. To this end we investigated whether a Drosophila melanogaster indirect flight muscle-specific promoter could function correctly in Culex quinquefascitaus since filarial worms molt and develop in the flight muscles of Culex mosquitoes prior to infecting a new vertebrate host. The Hermes transposable element-based transformation system was used to generate a transgenic line of Cx. quinquefasciatus. These transgenic mosquitoes express the green fluorescent protein (GFP) as a dominant marker placed under the control of the actin-88F (act88F) promoter of D. melanogaster. Genetic screens indicate that the transformation was successful, and that the marker inserted at a single locus. Significantly, the marker expression appears identical to that observed in D. melanogaster: GFP begins appearing in the developing flight muscles of the pupa and persists in the adult. The transgene was also shown to be sex-linked. Important implications of this research are: some promoters from D.melanogaster are functionally conserved in some mosquitoes; and act88F may be useful in driving expression of genes inhibitory to parasites that develop in mosquito flight muscles. Due to the insertion site of the transgene near the female sex locus, this particular transgenic line may also be utilized to clone female sex-specific genes.
Species 1: Diptera Culicidae Culex quinquefasciatus (southern house mosquito)
Keywords: transgenic, promoter
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