Cytochrome P450 genes: Molecular cloning and overexpression in a pyrethroid-resistant strain of Anopheles minimus mosquito

Abstract

We previously determined that physiological resistance in a laboratory-selected pyrethroid-resistant Anopheles minimus species A Theobald mosquito is associated with increased detoxification via a P450-mediated mechanism. A CYP6 gene, CYP6AA3, was subsequently cloned and found overexpressed in 2 resistant mosquito generations (F13 and F19). We report herein the cloning of CYP6P7 and CYP6P8 genes with full coding sequences from the same An. minimus mosquito colony strain. CYP6P7 and CYP6P8 encode proteins, each with 509 amino acids. CYP6P7 had the closest (81%) amino acid identity with Anopheles gambiae CYP6P2. CYP6P8 genes had 79% identity with An. gambiae CYP6P1. Using semiquantitative reverse transcription-polymerase chain reaction analysis, the mRNA expression level of CYP6P7 presented ∼2- and 4-fold increases in F19 and F25 deltamethrin-resistant populations, respectively, compared with the parent susceptible strain. CYP6P8 mRNA expression levels were not significantly different between the 3 filial generations. The overexpression of CYP6AA3 mRNA was greater than that of CYP6P7 in F19 and F25 resistant populations. The relative increase of both CYP6AA3 and CYP6P7 mRNA was correlated with increased resistance to deltamethrin in An. minimus. Copyright © 2005 by the American Mosquito Control Association, Inc.