Assembling the draft Reticulitermes flavipes CYPome to enhance the understanding of cytochrome P450 functions in relation to xenobiotic metabolism

Subterranean termites feed almost exclusively on wood lignocellulose which contains xenobiotics. Cytochrome P450 enzymes are one of the key mechanisms that termites likely use to metabolize these xenobiotics and minimize potential toxicity. Our lab has previously shown that approximately thirty 450 genes were up-regulated in R. flavipes with the consumption of wood (complex lignocellulose) relative to paper (cellulose). The objective of this work is to expand the knowledge of R. flavipes P450s by molecular characterization of candidate P450s and determining the tissue specificity. Approaches used in the present study included data mining for P450 genes from previous transcriptome sequencing, resequencing for verification, and determining tissue-dependent expression using RT-qPCR. Ten full-length P450 sequences were collected from prior microarray, pyrosequencing and illumina studies. The ten full-length sequences were homologous to CYP6, CYP15, CYP4, CYP302 and CYP9 families of other insect species.  Resequencing has further confirmed their identity to be P450s. All P450s studied contained the conserved heme binding domain (FXXGXXXCXG). Tissue specific expression of these P450s will be determined in salivary gland, midgut, hindgut, malphigian tubules and carcasses using RT-qPCR. We hypothesize that these R. flavipes P450s have specific tissue-associated expression profiles.