欧洲型舞毒蛾气味受体基因OrCo的克隆及序列分析

Molecular cloning and sequence analysis of the odorant receptor gene OrCo in Lymantria dispar(European gypsy moth)

【目的】克隆欧洲型舞毒蛾Lymantria dispar Linnaeus的OrCo气味受体基因,并分析其序列特征,为进一步研究舞毒蛾嗅觉机制提供有益参考。【方法】本研究利用RT-PCR和RACE方法,克隆获得欧洲型舞毒蛾OrCo受体基因cDNA全长序列,将该基因命名为LdisOrCo,在GenBank中的登录号为KF482409。【结果】序列分析结果显示,LdisOrCo开放阅读框全长为1 311 bp,编码436个氨基酸,序列中有7个跨膜区和高度保守的C端区域。序列联配分析表明,LdisOrCo基因的氨基酸序列与近缘种灰翅夜蛾(Spodoptera littoralis Or83b)的同源性高达89%,与鞘翅目台湾黑金龟(Holotrichia plumbea Or83b)同源性高达64%,与已经报道的其他昆虫的嗅觉受体同源性都在60%以上,特别是在C端几乎完全一致。【结论】嗅觉受体OrCo(Olfactory receptor coreceptor)在不同昆虫体内高度保守,克隆欧洲型舞毒蛾OrCo基因可以为进一步研究舞毒蛾气味受体的功能,OrCo基因的进化以及揭秘嗅觉机制奠定基础。

[Objectives] Exploring the mechanism of olfaetion in Lymantria dispra （European gypsy moth） by cloning and analyzing the olfactory receptor coreeeptor （OrCo） gene. [Methods] The full-length eDNA sequence of LdisOrCo was cloned using reverse transcription polymerase chain reaction （RT-PCR） and rapid amplification of eDNA ends （RACE） （GenBank accession number KF482409）. [Results] Sequence analysis revealed that the open reading frame （ORF） consisted of 1 311 nucleotides, encoding 436 amino acids with 7 putative transmembrane domains, and has a highly conserved C-terminus. Alignment analysis indicated that the nucleotide similarity of LdisOrCo with Or83b of Spodoptera littoralis and Holotrichiaplumbea was 89%, 64%, respectively, and that it shares high homologies （〉60%） with the corresponding receptor from other insects; the sequence of the C-terminus is almost the same. [Conclusion] The highly conserved olfactory receptor gene ofLymantria dispra （European gypsy moth） was sequenced in order to provide a basis for exploring the functions of the olfactory receptor, the evolution of the OrCo gene and the mechanism of insect olfaetion.