Ninety‐four putative G protein‐coupled receptors (GPCRs) were identified in the Musca domestica genome. They were annotated and compared with their homologues in Drosophila melanogaster. Phylogenetic analyses of the GPCRs from both species revealed that several family members shared a closer relationship based on the domain architecture. The expression profiles of these genes were examined by quantitative real‐time PCR amongst three strains of the house fly, a near‐isogenic line strain with imidacloprid resistance (N‐IRS), the corresponding susceptible strain (CSS) and another strain derived from field populations with imidacloprid resistance (IRS). We found that five GPCR genes were upregulated in the N‐IRS and eight GPCR genes were upregulated in the IRS strains compared to the CSS strain. The transgenic lines of D. melanogaster with the GPCR genes (LOC101899380 in the N‐IRS strain and LOC101895664 in the IRS strain) exhibited significantly increased tolerance to imidacloprid, and higher expression of cytochrome P450 genes. Bioinformatic analysis of LOC101899380 was carried out based on its full‐length nucleic acid sequence and putative amino acid sequence, and it was named Methuselah‐like10 (Mthl10) owing to its homology with D. melanogaster Mthl10. A cell‐base cell counting kit‐8 toxicity assay demonstrated that the expression of the GPCR gene LOC101899380 in Spodoptera frugiperda (Sf9) cells using a baculovirus‐mediated expression system can elevate the cell tolerance to imidacloprid, indirectly supporting the hypothesis that the GPCR gene LOC101899380 plays some role in imidacloprid resistance. These results should be useful for furthering understanding of the regulatory pathway by which house flies develop resistance.

中文翻译：

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G蛋白偶联受体相关基因在细胞色素P450介导的家蝇抗家蝇对双吡虫啉的抗性中的作用。

在家蝇基因组中鉴定出94个假定的G蛋白偶联受体（GPCR）。他们被注释并与果蝇中的同系物进行了比较。对这两个物种的GPCR的系统发育分析表明，基于域结构，几个家族成员有着更紧密的关系。这些基因的表达谱通过定量实时PCR检测了三只家蝇菌株，具有吡虫啉抗性的近等基因系菌株（N-IRS），相应的易感菌株（CSS）和另一种来自田间的菌株吡虫啉抗药性（IRS）的人群。我们发现，与CSS菌株相比，IRS菌株中有五个GPCR基因在N-IRS中被上调，而八个GPCR基因在IRS菌株中被上调。的转基因品系果蝇与GPCR基因（LOC101899380在N-IRS应变和LOC101895664在IRS菌株中）对吡虫啉的耐受性显着提高，并且细胞色素P450基因的表达更高。根据LOC101899380的全长核酸序列和推定的氨基酸序列进行了生物信息学分析，由于它与D. melanogaster Mthl10具有同源性，因此被命名为Methuselah-like10（Mthl10）。的细胞基细胞计数试剂盒-8毒性分析表明，GPCR基因的表达LOC101899380在草地贪夜蛾使用杆状病毒介导的表达系统（的Sf9）细胞可以提高细胞的耐受性吡虫啉，间接地支持这一假设的GPCR基因LOC101899380在吡虫啉抗性中起一定作用。这些结果应有助于进一步了解家蝇发展抗性的调控途径。