Molecular characterization of an arachnid sodium channel gene from the varroa mite (Varroa destructor).
Voltage-gated sodium channels are essential for the generation and propagation of action potentials in most excitable cells. They are the target sites of several classes of insecticides and acaricides. Isolation of full-length sodium channel cDNA is a critical and often difficult step toward an understanding of insecticide and acaricide resistance. We previously cloned and sequenced two overlapping cDNA clones covering segment 3 of domain II (IIS3) to segment 6 of domain IV (IVS6) of an arachnid sodium channel gene (named VmNa) from the varroa mite (Varroa destructor) (J. Apicultureal Res. 40 (2002) 5.). In this study, we isolated three more overlapping cDNA clones and revealed the entire coding region of VmNa (Genbank accession number: AY259834), thus providing the first complete cDNA sequence of an arachnid sodium channel gene. The composite VmNa cDNA contains 6645 nucleotides with an open reading frame encoding 2215 amino acids. The deduced amino acid sequence of VmNa shares a 51% overall identity with Drosophila Para and a 41% identity with the mammalian sodium channel alpha-subunit Na(v)1.2. All hallmarks of sodium channel proteins are conserved in the VmNa protein. Three optional exons and one retained intron were identified in VmNa. The precise position and size of only one exon is conserved in three insect sodium channel genes and mammalian Na(v)1.6 genes in human, mouse and fish, whereas the other three are novel. Interestingly, one of the novel exons is located in the C-terminus, where no alternative exons have been identified in any other sodium channel gene.
Wang
Ruiwu
R
Department of Entomology, Michigan State University, East Lansing, MI 48824, USA.
Huang
Zachary Y
ZY
Dong
Ke
K
D000595:Amino Acid Sequence; D000818:Animals; D004594:Electrophysiology; D005091:Exons; D015723:Gene Library; D008925:Mites / Q000235:genetics* / Q000502:physiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015222:Sodium Channels / Q000235:genetics* / Q000502:physiology*; D013778:Tetrazolium Salts
D015222:Sodium Channels; D013778:Tetrazolium Salts; C013574:iodonitrotetrazolium