DOI: 10.5061/dryad.v218g
title: Data from: Positional cloning of rp2 QTL associates the P450 genes CYP6Z1, CYP6Z3 and CYP6M7 with pyrethroid resistance in the malaria vector Anopheles funestus

description: Pyrethroid resistance in Anopheles funestus is threatening malaria control
 in Africa. Elucidation of underlying resistance mechanisms is crucial to
 improve the success of future control programs. A positional cloning
 approach was used to identify genes conferring resistance in the
 uncharacterised rp2 QTL previously detected in this vector using F6
 Advanced Intercross Lines (AIL). A 113 kb BAC clone spanning rp2 was
 identified and sequenced revealing a cluster of fifteen P450 genes and one
 salivary protein gene (SG7-2). Contrary to An. gambiae, AfCYP6M1 is
 triplicated in An. funestus while AgCYP6Z2 ortholog is absent. 565 new
 SNPs were identified for genetic mapping from rp2 P450s and other genes
 revealing high genetic polymorphisms with 1 SNP every 36bp. A significant
 genotype/phenotype association was detected for rp2 P450s but not for a
 cluster of cuticular protein genes previously associated with resistance
 in An. gambiae. QTL mapping using F6 AIL confirms the rp2 QTL with an
 increase logarithm of odds (LOD) score of 5. Multiplex gene expression
 profiling of 15 P450s and other genes around rp2 followed by individual
 validation using qRT-PCR indicated a significant over-expression in the
 resistant FUMOZ-R strain of the P450s AfCYP6Z1, AfCYP6Z3, AfCYP6M7 and the
 glutathione-s-transferase GSTe2 with respective fold-change of 11.2, 6.3,
 5.5 and 2.8. Polymorphisms analysis of AfCYP6Z1 and AfCYP6Z3 identified
 amino acid changes potentially associated with resistance further
 indicating that these genes are controlling the pyrethroid resistance
 explained by the rp2 QTL. The characterisation of this rp2 QTL
 significantly improves our understanding of resistance mechanisms in An.
 funestus.
description: Full sequence of rp2 BAC
 23 07 12Full length sequence of
 the 113kb BAC clone spanning the rp2 QTL. This sequence was assembled from
 6 contigs. It contains 15 P450 genes and the 1 salivary protein and is
 located on chromosome 2L.Genotype file for Family
 1 and 10 for QTL mappingGenotypes for different
 genetic markers in the two isofemale families (1 and 10) used for genetic
 and QTL mapping. Resistant individuals are coded with "1" while
 susceptible are coded with "0".DNA sequences for SNP
 identification 23 07 12DNA sequences from
 direct sequencing of different genes in F1 samples of the isofemale
 families Fam1 and Fam10. These sequences were used for SNP identification
 and genetic variability analysis using DnaSP program.CYP6Z1 cloned full
 length for FG FZ 23 07 12Full length sequences of
 the P450 gene CYP6Z1 between resistant (FZ for FUMOZ-R) and susceptible
 (FG for FANG). These sequences were obtained after
 cloning.CYP6Z3 cloned full
 length for FG FZ 23 07 12Full length sequences of
 the P450 gene CYP6Z3 between resistant (FZ for FUMOZ-R) and susceptible
 (FG for FANG). These sequences were obtained after
 cloning.qRTPCR data and PCR
 efficienciesqRT-PCR Ct values and
 PCR efficiencies for the genes tested between the resistant FUMOZ-R and
 the susceptible FANG strain.