title: Functional analysis of general odorant binding protein 2 from the meadow moth, Loxostege sticticalis L. (Lepidoptera: Pyralidae).
abstract: Odorant binding proteins play a crucial role in transporting semiochemicals across the sensillum lymph to olfactory receptors within the insect antennal sensilla. In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends. Recombinant LstiGOBP2 was expressed in Escherichia coli and purified by Ni ion affinity chromatography. Real-time PCR assays indicated that LstiGOBP2 mRNA is expressed mainly in adult antennae, with expression levels differing with developmental age. Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants. Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China. jyin@ippcaas.cn
authors.forename: Jiao
authors.initials: J
authors.lastname: Yin
authors.forename: Honglin
authors.initials: H
authors.lastname: Feng
authors.forename: Hongyan
authors.initials: H
authors.lastname: Sun
authors.forename: Jinghui
authors.initials: J
authors.lastname: Xi
authors.forename: Yazhong
authors.initials: Y
authors.lastname: Cao
authors.forename: Kebin
authors.initials: K
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057506:Arthropod Antennae / Q000502:physiology; D001483:Base Sequence; D004568:Electrodiagnosis / Q000379:methods; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D020869:Gene Expression Profiling; D008024:Ligands; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000145:classification / Q000235:genetics*; D016366:Open Reading Frames; D010802:Phylogeny; D011485:Protein Binding; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment
chemical_list: D005456:Fluorescent Dyes; D008024:Ligands; D018035:Receptors, Odorant; D011993:Recombinant Fusion Proteins; C045648:odorant-binding protein