为了解新发现的农业害虫双委夜蛾Athetis dissimilis(Hampson)非典型嗅觉受体Orco的分子特征与表达,通过分析转录组数据并利用RT-PCR技术克隆了双委夜蛾Orco基因,并采用实时定量PCR技术研究了该基因的组织表达谱。结果显示,双委夜蛾非典型嗅觉受体Orco基因开放阅读框全长1 422 bp,编码473个氨基酸,经氨基酸结构预测具有7个跨膜区,N端在膜内,C端在膜外;该基因编码的氨基酸序列与其它昆虫Orco序列相似性极高,因此将该基因命名为Adis Orco(Gen Bank登录号:KR632987),此氨基酸序列C末端第6跨膜区和第7跨膜区之间以及第7跨膜区的序列保守性最高;PCR结果显示,Adis Orco主要在成虫触角中表达,且在雄蛾触角中的表达量是雌蛾触角中的4.2倍,在足、翅、下唇须和喙等组织中也有少量表达。
In order to understand the molecular characteristics and expression pattern of olfactory receptor co-receptor( Orco) gene of Athetis dissimilis( Hampson),a new agricultural pest,the Orco was cloned through using transcriptome data analysis and RT-PCR techniques, and its expression pattern was analyzed in different tissues by real-time quantitative PCR method. The results showed that the Orco gene of A. dissimilis contained a 1 422-bp open reading frame( ORF) that encoded 473 amino acid residues,and the deduced Orco protein possessed structural characteristics of seven transmembrane domains,an intracellular N-terminus and an extracelluar C-terminus. The amino acid sequence of Orco gene had high similarity with the identified Orco from other insects; therefore,this gene was designated as Adis Orco( Gen Bank accession no. KR632987). The sequences between the sixth and seventh transmembrane domains,and the seventh transmembrane domains in the C-terminus showed the most conservative. The Adis Orco gene was highly expressed in antennae of adult A. dissimilis,and the expression level in adult males was 4. 2 times higher than that of the adult female,and low-level expression was also observed in labial palpi,proboscis,legs and wings.