FIGURE 1.

(A) Linear plasmid map showing the two genes (DsRed2 and tTAV) between the non-autonomous vector piggyBac sequences, inserted in the OX5034 Aedes aegypti strain. Due to the splicing module tTAV protein is only expressed in females in the absence of tetracycline family antibiotics. (B) Sex-specific splicing of endogenous Aeadsx and of the Aeadsx-ubiquitin-tTAV gene was confirmed in Aedes aegypti mosquitoes (pupal life stage, mosquitoes reared with doxycycline in the rearing medium) by RT-PCR reactions using either primers SS1997 and TD3349 (endogenous dsx), or TD226 and SJ124 (OX5034 dsx). All splicing isoforms of the endogenous Aeadsx gene (M, F1 and F2) were detected in WT as well as OX5034 mosquitoes in a sex-specific manner. The Aeadsx-ubiquitin-tTAV gene was also splicing in a sex-specific manner and was only detectable in mRNA extracted from OX5034 Aedes aegypti mosquitoes. In the absence of tetracycline antibiotics, the F2 isoform mRNA results in female-specific tTAV protein expression, and dsx and ubiquitin amino acids are cleaved from tTAV protein by endogenous cellular processes. Red octagons indicate stop codons.