title: Isolation and sequence analysis of P450 genes from a pyrethroid resistant colony of the major malaria vector Anopheles funestus.
abstract: Pyrethroid resistance has been demonstrated in populations of Anopheles funestus from South Africa and southern Mozambique. Resistance is associated with elevated P450 monooxygenase enzymes. In this study, degenerate primers based on conserved regions of Anopheles gambiae P450 CYP4, 6 and 9 families were used to amplify genomic and cDNA templates from A. funestus. A total of 12 CYP4, 12 CYP6 and 7 CYP9 partial genes have been isolated and sequenced. BLAST results revealed that A. funestus P450s generally have a high sequence identity to A. gambiae with above 75% identity at the amino acid level. The exception is CYP9J14. The A. gambiae P450 showing highest identity to CYP9J14 exhibits only 55% identity suggesting that CYP9J14 may have arisen from a recent duplication event. Molecular phylogenetic analysis based on amino acid sequences also supported this hypothesis. Intron positions, but not size, were highly conserved between the two species. The high level of orthology that exists in the P450 gene families of these two species may facilitate the prediction of individual P450 protein function.
authors.affiliation: Vector Control Reference Unit, National Institute for Communicable Diseases, Private bag X 4, Sandringham 2131, South Africa.
authors.forename: Dolphine A
authors.initials: DA
authors.lastname: Amenya
authors.forename: Lizette L
authors.initials: LL
authors.lastname: Koekemoer
authors.forename: Ashley
authors.initials: A
authors.lastname: Vaughan
authors.forename: John C
authors.initials: JC
authors.lastname: Morgan
authors.forename: Basil D
authors.initials: BD
authors.lastname: Brooke
authors.forename: Richard H
authors.initials: RH
authors.lastname: Hunt
authors.forename: Hilary
authors.initials: H
authors.lastname: Ranson
authors.forename: Janet
authors.initials: J
authors.lastname: Hemingway
authors.forename: Maureen
authors.initials: M
authors.lastname: Coetzee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000187:drug effects* / Q000235:genetics*; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D019143:Evolution, Molecular; D005091:Exons; D020440:Gene Duplication; D017344:Genes, Insect; D007305:Insecticide Resistance / Q000235:genetics; D007438:Introns; D008969:Molecular Sequence Data; D005810:Multigene Family; D010802:Phylogeny; D011722:Pyrethrins / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D018076:DNA, Complementary; D011722:Pyrethrins; D003577:Cytochrome P-450 Enzyme System