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Identification and Molecular Cloning of Putative Odorant-Binding Proteins and Chemosensory Protein from the Bethylid Wasp, <Emphasis Type="Italic">Scleroderma guani</Emphasis> Xiao et Wu
Authors:Daguang Lu  Xiangrui Li  Xiaoxia Liu  Qingwen Zhang
Affiliation:(1) Department of Entomology, College of Agriculture and Biotechnology, China Agricultural University, Beijing, 100094, China
Abstract:Two putative odorant-binding proteins (OBPs) and one putative chemosensory protein (CSP) from females of the ant-like bethylid wasp, Scleroderma guani Xiao et Wu (Hymenoptera: Bethylidae), were identified and cloned. The putative OBPs and CSP were identified by nondenaturing polyacrylamide gel electrophoresis (native-PAGE). 3′ rapid amplification of cDNA ends (3′RACE) was performed to obtain the sequences of the mature proteins by using degenerate primers designed from N-terminal sequences. Gene-specific primers for 5′ rapid amplification of cDNA ends (5′RACE) were designed according to 3′RACE results and used in polymerase chain reaction (PCR) to obtain full-length sequences. The proteins (Sgua-OBP1, Sgua-OBP2, and Sgua-CSP1) encode 133, 142, and 129 amino acid-deduced sequences, respectively. Prediction of signal peptide sequences matches the N-terminal amino acid sequence of the isolated proteins. Database searches suggest that the Sgua-OBP1 and Sgua-OBP2 are homologs of OBPs from other insects, and Sgua-CSP1 shares a high level of identity with previously described CSPs. Daguang Lu and Xiangrui Li contributed equally to this work
Keywords:Chemosensory protein  Hymenoptera  Molecular cloning  Nondenaturing polyacrylamide gel electrophoresis (PAGE)  Odorant-binding protein  Olfaction  Rapid amplification of cDNA ends (RACE) PCR            Scleroderma guani
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