Olfaction plays a crucial role in many aspects of insect behaviour, including host selection by agricultural pests and vectors of human disease. Insect odorant-binding proteins (OBPs) are thought to function as the first step in molecular recognition and the transport of semiochemicals. The whole genome sequence of the fruit fly Drosophila melanogaster has been completed and a large number of genes have been annotated as OBPs, based on the presence of six conserved cysteine residues and a conserved spacing between the cysteines. These proteins can be divided into three distinct subgroups; those with only one six-cysteine motif, those with two such motifs and those with one motif, three extra conserved cysteines and a conserved proline immediately after the sixth cysteine. This study concentrates on the last two subgroups, referred to as 'dimer' OBPs and 'Plus-C' OBPs, respectively. We determined the tissue-specific transcript levels of all of these OBP genes of D. melanogaster using semiquantitative RT-PCR. The results showed that the expression patterns can vary within a subgroup of genes and that this technique is valuable for assessing which of the putative OBP genes are likely to be involved in Drosophila olfaction. The publicly available genomes of another fruit fly Drosophila pseudoobscura, the malaria mosquito Anopheles gambiae and the yellow fever mosquito Aedes aegypti were searched by Blast against each Plus-C OBP and dimer OBP of D. melanogaster. Related genes were found in all of the other species and the relationships of these with the D. melanogaster genes and their possible biological functions are discussed.