Novel cytochrome P450s, CYP6BB1 and CYP6P10, from the salt marsh mosquito Aedes sollicitans (Walker) (Diptera: Culicidae)

Abstract

Based on the conserved heme-binding region and the charge pair consensus of insect cytochrome P450s, two novel full-length P450 cDNAs, CYP6BB1 and CYP6P10, were cloned from the salt marsh mosquito Aedes sollicitans (Walker). CYP6BB1 and CYP6P10 had open reading frames of 1,518 and 1,521 nucleotides encoding 506 and 507 amino acid residue proteins, respectively. Several alleles with amino acid substitutions were found both in CYP6BB1 and CYP6P10. The deduced proteins are typical microsomal P450s sharing signature sequences with other insect CYP6 P450s. Sequence analysis showed that both CYP6BB1 and CYP6P10 shared highest sequence identities with P450 CYP6P4, 56% and 65%, respectively. Phylogenetic analysis showed both CYP6BB1 and CYP6P10 were grouped into the clade containing several P450s from subfamily CYP6P. Real-time RT-PCR analysis showed CYP6BB1 but not CYP6P10 transcription in females was significantly increased 24 h after a blood meal. Neither CYP6BB1 nor CYP6P10 were life stage or gender specific. Protein expression experiments are needed to determine the functions of these proteins.

CYP9A12andCYP9A17in the cotton bollworm,Helicoverpa armigera: sequence similarity, expression profile and xenobiotic response

“…Therefore, identification and characterisation of new cytochrome P450 genes have become a very attractive research area. PCR‐based strategies (RT‐PCR, RACE) have been widely and effectively used in sequencing P450 genes 17, 30–32. One central consideration to this approach is the design of primers for PCR.…”

“…It is well known that P450 expression profiles are diverse in insects 37. For example, Cyp4d1 in Drosophila, CYP6A1 in the house fly and CYP6BB1 in A. sollictitans are expressed in all developmental stages,31, 38, 39 whereas CYP6Z1 in A. gambiae and CYP6D1 in Musca domestica L. are adult specific 40, 41 . CYP6L1 is specifically expressed in the reproductive tissues of adult male cockroaches,42 whereas CYP4G15 is predominantly expressed in the central nervous system of third‐instar larvae of Drosophila 43.…”

Molecular Cloning and Xenobiotic Induction of Seven Novel Cytochrome P450 Monooxygenases in Aedes albopictus

“…Degenerate primers: forward P450F (5′-GARACIYTIMGNAARTAYCC-3′) and reverse P450R (5′-ATRCADATICKIGGNCCYTC-3′) based on motif of family 6 P450 were used to generate partial sequence of family 6 P450 fragments ( Huang et al 2008 ). PCR was carried out with 94°C initial denaturation for 3 min, 40 cycles of 94°C for 30 s, 50°C for 30 s, and 72°C for 1 min, followed by a final extension at 72°C for 10 min.…”

Molecular characterization of two novel deltamethrin‐inducible P450 genes from Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae)

“…A different array of P450 isozymes exist in each insect species, even in different strains of the same species. A substrate molecule may be metabolized by different P450s, and the same P450 can metabolize different substrates (Huang et al, 2008). Recently, the availability of genome sequences in other insect species, such as An.…”