棉铃虫齿唇姬蜂嗅觉受体基因CchlOrco的克隆及组织表达谱分析

Cloning and tissue expression profiling of the olfactory receptor gene Cchl Orco in Campoletis chlorideae(Hymenoptera: Ichneumonidae)

【目的】昆虫的嗅觉受体(olfactory receptors,ORs)一般以气味分子特异的ORs与共受体(co-Receptor,Orco)通过形成异质二聚体在嗅觉感受中发挥关键作用,其中Orco由于具有序列的保守性而受到广泛的重视。本研究旨在克隆棉铃虫齿唇姬蜂Campoletis chlorideae的Orco基因,并对其组织表达谱进行分析。【方法】利用RT-PCR技术和转录组分析技术克隆棉铃虫齿唇姬蜂的Orco基因,并对其编码的氨基酸序列进行生物信息学分析;利用Realtime PCR技术对该基因在该蜂成虫不同组织中的表达量进行分析。【结果】获得了棉铃虫齿唇姬蜂Orco的全长c DNA序列,命名为Cchl Orco(Gen Bank登录号:KP255444)。序列分析表明,该基因开放阅读框全长1 437 bp,编码478个氨基酸,预测该氨基酸序列具有7个跨膜区。Cchl Orco主要在成虫触角中表达,且在雄蜂触角中的表达量最高,是雌蜂触角中表达量的8.0倍,而在其他组织中表达量极低。【结论】本研究克隆了棉铃虫齿唇姬蜂Cchl Orco序列全长,明确了其在成虫不同组织中的表达水平,为进一步研究该基因及其他嗅觉受体基因功能奠定了基础。

【Aim】Olfactory receptors（ ORs） interact with the co-receptor（ Orco） to form heterodimeric complexes that are critical for insect olfaction. Due to its highly conserved sequence,Orco has received extensive attention. The present study aims to clone and identify the Orco gene in Campoletis chlorideae,and analyze its tissue expression profiles. 【Methods】The Orco gene from C. chlorideae was cloned by RT-PCR and transcriptome technology, and the putative amino acid sequence was analyzed by bioinformatics methods. The expression level of Orco in different tissues of C. chlorideae adults was analyzed by real-time quantitative PCR. 【Results】The full-length c DNA sequence of Orco was obtained from C. chlorideae,and named as Cchl Orco（ Gen Bank accession no. KP255444）. Sequence analysis revealed that the open reading frame of Cchl Orco is 1 437 bp in length,encoding 478 amino acid residues with seven transmembrane domains. Real-time quantitative PCR results indicated that Cchl Orco was mainly expressed in the adult antennae of C. chlorideae,while expressed in other tissues at an extremely low level. The expression level of Cchl Orco in the antennae of male adults was 8. 0-fold as high as that in the antennae of female adults. 【Conclusion】The cloning of the full-length olfactory receptor gene Cchl Orco and its expression profiles provide a basis for future functional study of Cchl Orco and other olfactory receptor genes of C. chlorideae.