Available evidence has shown that transferrins are involved in iron metabolism, immunity and development in eukaryotic organisms including insects. Here we characterize the gene and message expression profile of Aedes aegypti transferrin 2 (AaTf2) in response to iron, bacterial challenge and life stage. We show that AaTf2 shares a low similarity with A. aegypti transferrin 1 (AaTf1), but higher similarity with mammalian transferrins and avian ovotransferrin. Iron-binding pocket analysis indicates that AaTf2 has residue substitutions of Y188F, T120S, and R124S in the N lobe, and Y517N, H585N, T452S, and R456T in the C lobe, which could alter or reduce iron-binding activity. In vivo studies of message expression reveal that AaTf2 message is expressed at higher levels in larva and pupa, as well as adult female ovaries 72h post blood meal (PBM) and support that AaTf2 could play a role in larval and pupal development and in late physiological events of the gonotrophic cycle. Bacterial challenge significantly increases AaTf1 expression in ovaries at 0 and 24h PBM, but decreases AaTf2 expression in ovaries at 72h PBM, suggesting that AaTf1 and AaTf2 play different roles in immunity of female adults during a gonotrophic cycle.