Figure 3.

Immunoblot analyses of tDefA peptide expression in transgenic mosquitoes after blood meal activation. (A) Protein extracts from individual 24 hPBM females of the host kh^w strain and the D1 transgenic line were tested by using polyclonal antibody to Ae. aegypti DefA and mAb to Ae. aegypti vitellogenin small subunit (27). Expression of the 4-kDa DefA peptide was observed in transgenic mosquitoes but not in the kh^w host strain. The expression of 66-kDa Vg-small subunit (Vg-S), used as a positive control for blood meal activation, was present at the same level in both the transgenic line and the host strain. (B) Immunoblot analysis of tissue- and sex-specific expression of tDefA peptide in transgenic mosquitoes. Protein extracts from hemolymph (H), fat bodies (FB), ovary (OV), carcass (C), 24 hPBM whole mosquito female, (F), and mosquito male (M) were analyzed. (C) Time course of tDefA protein accumulation in transgenic mosquitoes during the vitellogenic cycle after a single blood feeding. Previtellogenic female (PV), vitellogenic females 1 (1d), 3 (3d), and 22 (22d) days PBM and the hemolymph collected from 22-day-old female mosquitoes (H22d) were tested using DefA-specific antibodies. One mosquito-equivalent was loaded in each lane, except for the hemolymph sample from 22-day-old females, in which a four-mosquito equivalent was used.