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.