Molecular cloning and characterization of a novel P450 gene encoding CYP6BK18 from dastarcus helophoroides (Coleoptera: Bothrideridae)

Abstract

A novel cDNA clone encoding a cytochrome P450 gene, named CYP6BK18 (GenBank KC683905), was isolated by reverse transcription PCR from Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae), a natural enemy of beetles. The full-length cDNA sequence is 1,659 bp, containing a 1,533 bp open reading frame predicting a 510-amino acid protein possessing a transmembrane domain with a calculated molecular weight of 59.4 kDa and a theoretical pI of 8.94. The deduced amino acid sequence of CYP6BK18 showed a 59% identity with CYP6BK17 (GenBank XP-970481.1) from Tribolium castaneum. Phylogenetic analysis indicated that CYP6BK18 was most closely related to CYP6BK17 and CYP6BK14 (GenBank EFA05731.1) from T. castaneum. Expression patterns of CYP6BK18 in different tissues (head, oviduct, midgut, fat bodies, and Malpighian tubules), developmental stages (first-to sixth-instar larvae and adult) and 10 age groups of adult were analyzed by real-time quantitative PCR (RT-qPCR). The results showed that CYP6BK18 was highly expressed in adulthood. Also, RT-qPCR analysis among different age groups of adult showed that CYP6BK18 transcripts were abundant in the spawning period and peaked at the early stage of the adult development. Moreover, the tissue-specific expression levels of CYP6BK18 were in the order of midgut, Malpighian tubes and fatty body from high to low. These results suggest that cytochrome P450 CYP6BK18 may play a role in regulating the development and aging of D. helophoroides.