Fig. 4

Generation, characterization and Plasmodium gallinaceum infections of pBac[3XP3-eGFP-AeVg-PgpNT] transgenic mosquitoes. a EGFP fluorescence in both eyes of larvae, pupae and adults of transgenic mosquitoes (T) but not in wild-type controls (WT). Only in larvae EGFP could be detected in the neural tube. b Genomic DNA was extracted from transgenic lines pBac-PgpNT P#1, P#4 and F#8 and used for PCR with specific primers for the PgpNT fragment. c Detection of transgenic transcript from pBac-PgpNT P#1 mosquitoes. Total RNA was extracted from non-blood fed (NBF), 24, 48 and 72 h post-blood meal females mosquitoes and used on a RT-PCR reaction with specific primers for PgpNT sequence. The same RNA samples were also analysed by vitellogenin (Vg) and actin (Act) gene-specific primers (Additional file 1). d Transgenic pBac-PgpNT P#1 and wild-type adult females were fed in P. gallinaceum-infected chickens, five days later a second blood meal with healthy mice was offered. Salivary glands were dissected eight days after infected blood meal and sporozoite counted using phase-contrast microscopy. Solid circles and squares represent individual wild-type and transgenic mosquitoes respectively, with sporozoites detected in their salivary glands. The horizontal bars represent the median, a p value of <0.05 was considered statistically significant