【目的】分离和克隆昆虫触角中一类非常保守的嗅觉受体蛋白基因全序列。【方法】采用简并引物反转录多聚酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)技术克隆基因,用生物信息学方法对获得的cDNA全序列及推导的氨基酸序列进行分析。【结果】从甜菜夜蛾(Spodoptera exigua Huebner)和斜纹夜蛾[Spodoptera litufa(Fabricius)】雄蛾触角中扩增得到2个分别为1906bp和2483bp的OR83类化感蛋白基因的cDNA,开放阅读框(ORF)编码473个氨基酸,命名为SexiOR2和S1itOR2。通过在GenBank中进行序列的同源性比较,发现这2个鳞翅目化感蛋白新序列与已知蛾类昆虫的OR83b类化感蛋白氨基酸序列具较高的同源性。【结论】明确了所获得的2个鳞翅目蛋白新序列属于OR83b类化感蛋白。
[Objective] The aim of this study was to isolate the cDNA full sequence of an unusually highly conserved olfactory receptor (orthologue to the Drosophila melanogaster DOR83b) gene from the antennae of insects. [Method] The olfactory receptor genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. Bioinformatics methods were used to further analyze cDNA sequence obtained and putative amino acid sequence. [Result] Two cDNA full sequences of olfactory receptor (OR) gene were amplified from the antennae of male Spodoptera exigua and S. litura, and they were named SexiOR2 and SlitOR2. SexiOR2 and SlitOR2 contained 1 906 bp and 2 483 bp, and with deduced amino acid sequences of 473 residues. The sequence analysis indicated that the deduced amino acid sequences of the cDNA sequences shared high identity with OR83b orthologue chemoreceptor sequences from other previously reported moths. [ Conclusion ] It is inferred that the sequences obtained are cDNA sequences of OR83b orthologue chemoreceptor gene.