Tissue Expression Profile and Ligand Binding Affinity of the Chemosensory Protein Gene CpalCSP3 in Chrysopa pallens(Rambur)

Wang Sibao, Zhang Shuai, Luo Junyu, Li Zhaoqun, Lü Limin, Wang Chunyi, Cui Jinjie, Dong Shuanglin

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Cotton Science ›› 2015, Vol. 27 ›› Issue (3) : 260-267. DOI: 10.11963/issn.1002-7807.201503010

Tissue Expression Profile and Ligand Binding Affinity of the Chemosensory Protein Gene CpalCSP3 in Chrysopa pallens(Rambur)

  • Wang Sibao1,2, Zhang Shuai2, Luo Junyu2, Li Zhaoqun1,2, Lü Limin2, Wang Chunyi2, Cui Jinjie2*, Dong Shuanglin1*
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Abstract

Insect chemosensory proteins(CSPs) play important roles in olfactory perception by transporting hydrophobic chemical volatiles across the sensillum lymph to olfactory receptors, but this role in Chrysopa pallens(Rambur) has not been addressed. In this paper, a CSP gene CpalCSP3 was cloned and the tissue expression profile of the CpalCSP3 gene in male and female adults was determined by real-time quantitative polymerase chain reaction. Results showed that the CpalCSP3 gene was highly expressed in the heads and antennae of both sexes, suggesting its involvement in olfaction and gustation. To further explore the olfactory role of the gene, recombinant CpalCSP3 protein was obtained by the Escherichia coli expression system, and the binding affinities of CpalCSP3 protein for 84 plant volatiles were measured using fluorescent competitive binding assay. Among the tested compounds, nerolidol, (+)-cedrol and beta-ionone had high binding affinities to CpalCSP3 protein, with Ki values ranging from 8.79 to 9.79 μmol·L-1. In addition, phlorizin dihydrate, dodecyl aldehyde, geranyl acetate and (+/?)-alpha-pinene also showed high binding affinities, with Ki values from 11.54 to 19.06 μmol·L-1. Together, these results suggest that CpalCSP3 protein may play a role in olfaction of these plant volatiles, which facilitate host-plant searching in adult C. pallens.

Keywords

Chrysopa pallens / chemosensory protein / tissue expression profile / prokaryotic expression / fluorescence competitive binding assay

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Wang Sibao, Zhang Shuai, Luo Junyu, Li Zhaoqun, Lü Limin, Wang Chunyi, Cui Jinjie, Dong Shuanglin. Tissue Expression Profile and Ligand Binding Affinity of the Chemosensory Protein Gene CpalCSP3 in Chrysopa pallens(Rambur)[J]. Cotton Science, 2015, 27(3): 260-267. https://doi.org/10.11963/issn.1002-7807.201503010

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