Identification and Characterization of Two General Odorant Binding Protein Genes from the Oriental Fruit Moth, Grapholita molesta (Busck)

Abstract

Two novel general odorant-binding protein (GOBP) cDNAs (GmolGOBP1 and GmolGOBP2) were cloned and characterized from female antennal tissue of the oriental fruit moth, Grapholita molesta. We focused our investigation on this olfactory protein family by using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends-PCR (RACE-PCR). The full-length open reading frames of GmolGOBP1 and GmolGOBP2 were 492 and 483 nucleotides long, which encode 164 and 161 amino acid residue peptides, respectively. Protein signature analyses revealed that they each contained six conserved cysteines with an N-terminal signal sequence of 20 amino acids. The alignment of the two deduced protein sequences with other Lepidoptera GOBPs showed high sequence similarity (70-80%) with other full-length sequences from GenBank. Sequence similarity between the two GOBPs was only 48%, suggesting that the two proteins belong to different classes of lepidopteran GOBPs. RT-PCR analysis revealed that the two GOBP genes were expressed only in antennae of both sexes. Real-time PCR analysis further indicated that the transcript level of GmolGOBP1 was higher in males than in females, whereas the transcript level of GmolGOBP2 was higher in females than in males. Temporally, the two GOBP genes were expressed during the complete photoperiod (15L:9D). The highest transcript levels of GmolGOBP1 in both sexes and GmolGOBP2 in females were detected at the end of photophase and during scotophase. The expression of GmolGOBP2 in males remained at similar levels during the complete photoperiod. Based on these results, the possible physiological functions of GmolGOBPs are discussed.