, 1992). This approach has limitations as orthologs may be involved only in the detection of common this website ligands, and the chemical ecology of the malaria and the Southern house mosquitoes differ. For the current study we selected putative Cx. quinquefasciatus ORs from six phylogenetic groups, five of which with no An. gambiae orthologs. Following cloning, quantitative PCR analysis was performed to confirm expression in female antennae, and then the ORs were co-expressed with the obligatory co-receptor Orco in Xenopus oocytes for de-orphanization. As reported here, we have identified one OR that responds to multiple compounds and another that did not

respond to any compound http://www.selleckchem.com/products/wortmannin.html tested, in addition to an OR displaying stronger responses to plant-derived, natural mosquito repellents, and another sensitive to phenolic compounds, particularly eugenol. Amino acid sequences of mosquito ORs were combined to create an entry file for phylogenetic analysis in Mega 5.05 (Tamura et al., 2011). An unrooted consensus neighbor joining tree was calculated at default settings with pairwise gap deletions. Branch support was assessed by bootstrap analysis based on 1000 replicates. Seventy-six

An. gambiae, 99 Aedesc and 130 Cx. quinquefasciatus ORs were included in this analysis. Sequence alignments were performed with ClustalW2 (http://www.ebi.ac.uk/Tools/msa/clustalw2/). Sequences available in databases were screened for full-length functional ORs based on multiple alignments and prediction of transmembranes. Partial sequences, truncated sequences, and pseudogenes, based on current OR genes annotations, were omitted (AgamOR81; AaegOR6, 12, 18, 22, 29, 32, 35, 38, 39, 51, 54, 57, 64, 68, 73, 77, 82, 83, 86, 91, 97, 108, 112, 116, 118, 120,

126, 127, 128, 129, 130, 131; CquiOR3, 8, 9, 15, 17, 19, 26, 31, 33, 34, 35, 41, 49, 59, 66, 74, 76, 94, 100, 101, 102, 103, 104, 105, 111, 119, 124, 125, 129, 133, 134, 135, 138, 139, 140, 144, 147, 152, Niclosamide 158, 159, 160, 167, 168, 170, 172, 174, 176, 177, 178, 179, 180). Cx. quinquefasciatus mosquitoes used in this study were from a laboratory colony maintained at UC Davis. This colony was initiated with adult mosquitoes from a colony maintained by A.J.C. at the Kearney Agricultural Center, University of California, and started from mosquitoes collected in Merced, CA in the 1950s. In Davis, mosquitoes were kept in an insectary at 27 ± 1 °C, under a photoperiod of 16:8 h (L:D) for the last 3 years. Total RNA was extracted from one thousand 1–5-day-old female Cx. quinquefasciatus antennae with TRIzol reagent (Invitrogen, Carlsbad, CA). Antennal cDNA was synthesized from 1 μg of antennal total RNA using SMARTer™ RACE cDNA amplification kit according to manufacturer’s instructions (Clontech, Mountain View, CA).