title: Transcriptome Analysis of an Insecticide Resistant Housefly Strain: Insights about SNPs and Regulatory Elements in Cytochrome P450 Genes.
abstract: BACKGROUND
  Insecticide resistance in the housefly, Musca domestica, has been investigated for more than 60 years. It will enter a new era after the recent publication of the housefly genome and the development of multiple next generation sequencing technologies. The genetic background of the xenobiotic response can now be investigated in greater detail. Here, we investigate the 454-pyrosequencing transcriptome of the spinosad-resistant 791spin strain in relation to the housefly genome with focus on P450 genes.
  
  
  RESULTS
  The de novo assembly of clean reads gave 35,834 contigs consisting of 21,780 sequences of the spinosad resistant strain. The 3,648 sequences were annotated with an enzyme code EC number and were mapped to 124 KEGG pathways with metabolic processes as most highly represented pathway. One hundred and twenty contigs were annotated as P450s covering 44 different P450 genes of housefly. Eight differentially expressed P450s genes were identified and investigated for SNPs, CpG islands and common regulatory motifs in promoter and coding regions. Functional annotation clustering of metabolic related genes and motif analysis of P450s revealed their association with epigenetic, transcription and gene expression related functions. The sequence variation analysis resulted in 12 SNPs and eight of them found in cyp6d1. There is variation in location, size and frequency of CpG islands and specific motifs were also identified in these P450s. Moreover, identified motifs were associated to GO terms and transcription factors using bioinformatic tools.
  
  
  CONCLUSIONS
  Transcriptome data of a spinosad resistant strain provide together with genome data fundamental support for future research to understand evolution of resistance in houseflies. Here, we report for the first time the SNPs, CpG islands and common regulatory motifs in differentially expressed P450s. Taken together our findings will serve as a stepping stone to advance understanding of the mechanism and role of P450s in xenobiotic detoxification.
authors.affiliation: Department of Agroecology, Aarhus University, Slagelse, Denmark.
authors.forename: Khalid
authors.initials: K
authors.lastname: Mahmood
authors.affiliation: Department of Agroecology, Aarhus University, Slagelse, Denmark.
authors.forename: Dorte H
authors.initials: DH
authors.lastname: Højland
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Slagelse, Denmark.
authors.forename: Torben
authors.initials: T
authors.lastname: Asp
authors.affiliation: Department of Agroecology, Aarhus University, Slagelse, Denmark.
authors.forename: Michael
authors.initials: M
authors.lastname: Kristensen
mesh_terms: D000818:Animals; D016000:Cluster Analysis; D018899:CpG Islands / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000145:classification / Q000235:genetics; D004338:Drug Combinations; D020869:Gene Expression Profiling / Q000379:methods; D063990:Gene Ontology; D006793:Houseflies / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics; D007305:Insecticide Resistance / Q000235:genetics*; D007306:Insecticides / Q000494:pharmacology; D018942:Macrolides / Q000494:pharmacology*; D016366:Open Reading Frames / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D004338:Drug Combinations; D019476:Insect Proteins; D007306:Insecticides; D018942:Macrolides; D003577:Cytochrome P-450 Enzyme System; C415329:spinosad