Cloning and sequencing the calcitonin receptor-like receptor 3 (AaegGPRCal3) from female mosquitoes of Aedes aegypti (L.) (Diptera: Culicidae)

The biological role of calcitonin receptor-like receptors that belong to the family B of G-protein coupled receptors (GPCRs), in maintenance of water balance of females of mosquito Aedes aegypti is being studied. In Anopheles gambiae, the Angoga-DH₃₁ receptor (a calcitonin receptor-like receptor, AaegGPRCal1), functions in maintenance of water balance, as it is responsible for sodium excretion in Malpighian tubules post blood feeding. The presence of receptors involved in water balance has also been confirmed in the central nervous system and other peripheral tissues in insects. This project focuses on the Calcitonin 3 (AaegGPRCal3) GPCR, another member of family B. Cloning of the Cal3 GPCR was attempted from RLM cDNA synthesized from various tissues dissected from females of A.aegypti. PCR amplified DNA fragments were cloned and sequenced. Alignment of these sequences with the predicted genomic sequence (gene ID: AAEL009024; vectorbase.org) showed an exact match of 289 amino acids in the middle of the ORF, but a different sequence than the genomic prediction at the ORF 5’ end and an additional 36 amino acids at the ORF 3’ end. These results demonstrate that experimental validation of predicted sequences is necessary before attempting other genetic studies of these important GPCRs.