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 |
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Authors: | Daguang Lu Xiangrui Li Xiaoxia Liu Qingwen Zhang |
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Affiliation: | (1) Department of Entomology, College of Agriculture and Biotechnology, China Agricultural University, Beijing, 100094, China |
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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 |
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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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