(A) Schematic genome map of wBm shown as a circle. The relative locations (shown as closed arrows) of operon 1 (virB8-1, virB9-1, virB10, virB11, virD4, wsp) and the upstream gene ribA, operon 2 (virB3, virB4-1, virB6-1, virB6-2, virB6-3, and virB6-4), virB4-2, virB8-2, virB9-2, wBmxR1, and wBmxR2 are shown. (B) Expression of Wolbachia virB8-1 in various developmental stages of B. malayi. RNA from adult female, male, microfilaria, L3 and L4 stages were analyzed by quantitative RT-PCR. The relative gene expression of virB8-1 is represented relative to Wolbachia 16S rRNA. Data obtained from triplicate samples are expressed as a mean ± standard deviation. (C) Detection of VirB8-1 in protein lysates of adult female worms using anti-VirB8-1 antibody in Western blot analysis. Lane 1, SDS-PAGE gel showing purified recombinant VirB8-1 with an N-terminal His tag and deleted trans-membrane domain (rHis-ΔTM VirB8-1). Western blot analysis using rabbit serum generated against rHis-ΔTM VirB8-1 (lanes 2–5). Lane 2, lysate of E. coli expressing rHis-ΔTM VirB8-1; Lane 3, recombinant full length VirB8-1 with N-terminal His tag (rHis-VirB8-1); Lane 4, recombinant full length VirB8-1 minus tag (rVirB8-1); Lane 5, lysate of female B. malayi worms. Arrows indicate the band corresponding to the VirB8-1 protein.