双委夜蛾非典型嗅觉受体Orco的克隆、分子特征及表达
Cloning, molecular characteristics and expression pattern of the olfactory receptor co-receptor gene of Athetis dissimilis

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1422 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AdisOrco(GenBank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,AdisOrco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 AdisOrco (GenBank 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 AdisOrco 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.