The class I glutathione S-transferases (GSTs) of Anopheles gambiae are encoded by a complex gene family. We describe the genomic organization of three members of this family, which are sequentially arranged on the chromosome in divergent orientations. One of these genes, aggst1-2, is intronless and has been described. In contrast, the two A. gambiae GST genes (aggst1alpha and aggst1beta) reported within are interrupted by introns. The gene aggst1alpha contains five coding exons that are alternatively spliced to produce four mature GST transcripts, each of which contains a common 5' exon encoding the N termini of the GST protein spliced to one of four distinct 3' exons encoding the carboxyl termini. All four of the alternative transcripts of aggst1alpha are expressed in A. gambiae larvae, pupae, and adults. We report on the involvement of alternative RNA splicing in generating multiple functional GST transcripts. A cDNA from the aggst1beta gene was detected in adult mosquitoes, demonstrating that this GST gene is actively transcribed. The percentage similarity of the six cDNAs transcribed from the three GST genes range from 49.5% to 83.1% at the nucleotide level.