Transfection ofWolbachia pipientisintoDrosophilaEmbryos

Wolbachia is a genus of obligate intracellular Alpha-Proteobacteria represented by the type species Wolbachia pipientis (Dumler et al., 2001). Wolbachia commonly reside within cytoplasmic vacuoles of arthropods and helminths (Werren and Windsor, 2000; Casiraghi et al., 2004); vertebrate infections have not been identified. Wolbachia are maternally transmitted from mothers to offspring though the embryonic cytoplasm. Wolbachia are able to induce a diverse range of phenotypes in their invertebrate hosts, ranging from classical mutualism to reproductive parasitism. Examples of the latter include male killing, host feminization, parthenogenesis, and cytoplasmic incompatibility (reviewed in Dobson, 2003a). Current Wolbachia research foci include examining the impacts of Wolbachia infection on host evolution, characterizing the mechanisms by which Wolbachia manipulate invertebrate hosts, and developing applied strategies that employ Wolbachia for pest and disease control. Wolbachia transfection has proven a useful technique for addressing questions within each of these research foci. This unit describes a method for Wolbachia transfection via embryonic microinjection.