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Cloning and sequence characteristics analysis of nicotinic acetylcholine receptor α4 subunit gene fromTetranychus cinnabarinus

Nicotinic acetylcholine receptor(nAChR)was the gated ion channel response to acetylcholine and one of the most important insecticide targets.In this study,the nAChRα4subunit gene of Tetranychus cinnabarinus(Boisduval)was cloned by RT-PCR and RACE.The cDNA had a length of 2 023 bp,and its Open Reading Frame(ORF)had a length of 1 635 bp encoded a protein of 544 amino acid residues.Molecular weight(Mr)of the protein is 62.20 kDa,and the isoelectric point(pI)is5.00.It was named as Tcα4(GenBank accession:No.KP694228)according to its amino acid homology to those of T.urticae and other insects.The nAChRα4subunit of T.cinnabarinus had rypical character of nAChR units with a N-terminal transmembrane domain and a C-terminal extracellular domain.Tcα4had a homology of 98.71%to T.urticae nAChRα4,and only20.07%~43.16% homology to other nAChRs of T.urticae;and had a range of 45.96%~56.07% homology to other insectsnAChRα4subunits.Phylogenetic tree analysis showed that Tcα4,T.urticae and Ixodes scapularis nAChRα4were classified in the same cluster,and were far from the others.The cloning and character analysis of Tcα4lay a solid foundation for the study of other nAChR subunits fromT.cinnabarinus and development of new biological miticide targeted to nAChR.