Molecular characterization of two nicotinic acetylcholine receptor subunits from Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae).

Two nicotinic acetylcholine receptor (nAChR) subunit genes, Lbalpha1 and Lbalpha8, were isolated and characterized from psocid, Liposcelis bostrychophila Badonnel, using the rapid amplification of cDNA ends (RACE) technique. They are the first two nAChR family members isolated from the insect order of Psocoptera. The full-length cDNAs of Lbalpha1 (GenBank accession number: EU871527) and Lbalpha8 (EU871526) consist of 2,025 and 1,763 nucleotides, respectively, and an open reading frame of 1,644 and 1,608 bp encoding 547 and 535 amino acid proteins, respectively. Both genes have typical features of nAChR family members, though they share only 56% identity in amino acid sequence. The dendrogram generated by the MEGA 3.1 program shows that the protein deduced by Lbalpha1 had the closest phylogenetic relationship to Agamalpha1 from Anopheles gambiae and Amelalpha1 from Apis mellifera, and Lbalpha8 shares the highest identity with Agamalpha8 from An. gambiae and Amelalpha8 from A. mellifera. Quantitative real-time PCR analysis showed that Lbalpha1 was expressed 2.03-6.54-fold higher than Lbalpha8 at the different developmental stages of L. bostrychophila. The highest expression levels of Lbalpha1 and Lbalpha8 were both detected at adult stage and the lowest were at the third and fourth nymphal stages, respectively. There was a stable and relatively low expression level for Lbalpha1, whereas there was a descending expression pattern for Lbalpha8 in the 1st through the 4th nymphal stadia.