Cytochrome P450 Genes Expressed in Phasmatodea Midguts

In June 2021, representative sequences for insect cytochrome P450s were downloaded from NCBI, limiting the search to those in the UniProtKB database. The resulting 111 sequences were used as a query to mine the above transcriptomes using tblastn with an expect value threshold of e-10. These were manually annotated by removing truncated sequences, using the ExPASy online translation tool to obtain the complete amino acid sequences, removing duplicates using the sRNAtoolbox webserver, and confirming that the sequences were cytochrome P450s by identifying them using blastp against the NCBI database. The resulting sequences were combined with the representatives from NCBI, aligned using the Clustal W program built into the software MEGA version X. Any sequences missing the heme-binding domain FXXGXXXCXG/A, which is a signature motif for CYPs, were deleted. The sequences were also checked for the presence and absence of other four signature motifs from insect CYPs: helix C (WxxxR), helix I (GxE/DTT/S), helix K (ExLR), and PERF (PxxFxPE/DRE/F).