Cloning and sequence analysis of cDNA encoding aquaporin (AQP) gene from Anopheles sinensis

Objective To clone and analyze the full-length sequence of aquaporin gene of Anopheles sinensis(AsAQP),so as to provide an insight into its biology functions.Methods The degenerate primers were used to amplify conserved region of AQP from An.sinensis cDNA.After then,the full-length cDNA of AsAQP was obtained by rapid amplification of cDNA ends(RACE).Concurrently,the bioinformatics methods were applied to analyze the obtained sequence.Results The obtained full-length cDNA of AsAQP consisted of 762 bp and 253 deduced amino acids with a predicted molecular mass of 63.2 kD.Bioinformatics analysis demonstrated that AsAQP had a typical structure with six membrane-spanning domains and an internal symmetry showing two highly conserved Asn-Pro-Ala(NPA) motif and possessing the consensus sequence of major intrinsic protein(MIP)superfamily.The AsAQP shared the identities of 76% and 78% with those of Culex quinquefasciatus and Aedes aegypti AQPs,respectively.Phylogenetic analysis indicated that AsAQP was clustered with Aedes and Culex AQPs.Conclusions The full-length AsAQP is cloned by degenerate primers and RACE from An.sinensis.The AsAQP gene is a member of MIP protein family,and has the typical function region.The study lays the foundation for further research on the function of AsAQP.