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本研究构建夜蛾科昆虫化学感受蛋白系统进化树,发现化学感受蛋白在种间保守性强,并与气味结合蛋白明显分为两类。运用反转录RT-PCR方法克隆了Sexi CSP3基因,并在大肠杆菌中正确表达,重组Sexi CSP3蛋白纯化后通过荧光竞争结合实验测定其与24种外源气味分子的结合特性。结果表明:Sexi CSP3与苯甲醛、苯乙酮、2-戊基-3-苯丙基-烯醛、1-苯基-1-丁酮、对甲氧基苯甲醛的结合能力较强,解离常数为0.76、1.39、2.23、3.23和5.30μmol/L,推测Sexi CSP3可能参与了甜菜夜蛾对这5种化学物质的识别过程。
In this study, we constructed the phylogenetic tree of the insects of the noctuid insects and found that the chemosensitive proteins are highly conserved among species and clearly classified into two types with odor-binding proteins. The Sexi CSP3 gene was cloned by RT-PCR and correctly expressed in E. coli. The recombinant Sexi CSP3 protein was purified and its binding properties to 24 exogenous odorant molecules were determined by fluorescence competition-binding assay. The results showed that the binding ability of Sexi CSP3 to benzaldehyde, acetophenone, 2-pentyl-3-phenylpropyl-enal, 1-phenyl-1-butanone and p-methoxybenzaldehyde was strong, From the constants 0.76, 1.39, 2.23, 3.23 and 5.30 μmol / L, it is speculated that Sexi CSP3 may be involved in the identification of these five chemicals by S. exigua.