Analysis of differentially expressed genes is a common molecular biological tool to investigate changes in mosquito genes after a bloodmeal or parasite exposure. We report here the characterization of a differentially expressed actin gene, CNAct-1, from the subtropical mosquito, Culex nigripalpus Theobald (Diptera: Culicidae). The CNAct-1 genomic clone is 1.525 kb, includes one 66-bp intron, and a 328-bp 3′-untranslated region. The 376-amino acid putative translation product shares high similarity with muscle-specific actin proteins from other insects, including Culex pipiens pipiens L., Aedes aegypti (L.), Anopheles gambiae Giles and Drosophila melanogaster (Meigen). CNAct-1 is expressed in second and third instars, late pupae, and adult females and males. Interestingly, Cx. nigripalpus actin was highly expressed in female mosquito midgut tissue isolated 6–12 h after ingestion of a bloodmeal. This expression profile indicates a unique function for CNAct-1 in midgut processes that are initiated after blood ingestion.