Kinetic parameter of <i>Ae. aegypti</i> recombinant ALDH isoforms.

<p>Kinetic studies were performed by varying the concentration of PBald and cofactor NAD⁺ at fixed saturated concentrations of NAD⁺ and PBald, respectively. The oxidation of PBald to PBacid was monitored by the formation of NADH in the reaction at 37°C for 4 min. Three independent assays were performed. The results are shown as the mean ± SE.</p

Specific activity of <i>Ae.aegypti</i> recombinant ALDH isoforms to oxidise PBald.

<p>Recombinant ALDH (5 µg) was incubated with 2 mM PBald in the presence of 3 mM NAD+ in 0.1 M Tris-Cl buffer, pH 7.4 at 37°C for 10 min. PBacid formation was determined by HPLC as described. Three independent assays were performed. The results are shown as the mean ± SE.</p

Substrate specificity of <i>Ae. aegypti</i> recombinant ALDH isoforms.

<p>ALDH activity was performed in the presence 4 mM PBald and 2.5 mM NAD(P)⁺. The oxidation of PBald was monitored by the formation of NAD(P)H.</p

Historical demographic analyses.

<p>(A) Mismatch distribution computed using the software Arlequin 3.1. Histograms show the observed distribution; lines show the expected distribution under a model of sudden population expansion. (B) Bayesian Skyline Plot, constructed using the software beast 1.6.1. Population size (<i>y</i>-axis) is measured as the product of effective population size per generation length (<i>N</i>_eτ). The solid line is the median estimate, and the grey areas show the 95% Highest Posterior Density (HPD) limits. Time (<b><i>x</i></b> axis) is expressed in years before present (BP).</p

Deduced amino acid sequences of <i>Ae. aegypti</i> ALDH9948 and ALDH14080.

<p>Sequences shown are from the PMD-R strain. The amino acid sequences were aligned using ClustalW. Letters in bold indicate 100% conservation between the 3 sequences. Dashes are used to denote gaps introduced for maximum alignment.</p

Transcription profiles of <i>ALDH9029</i>, <i>ALDH9948</i> and <i>ALDH14080</i> in three strains of <i>Ae. aegypti</i>.

<p>Complementary DNA from three different biological replicates (ten mosquitoes each) was used as templates. Four life-stages were analysed: larvae (L), pupae (P), adult male (M), and adult female (F). Each sample was analysed in duplicate in each experiment, and the results were averaged from three independent experiments. The mRNA copy numbers were determined by comparison with known concentrations of standard plasmids and normalised against the copy number of the ribosomal S7 transcript. Error bars indicate standard error of the mean. Statistically significant differences were evaluated with ANOVA followed by Tukey's multiple comparison test (*<i>p<</i>0.05 versus New Orleans strain; ^#<i>p</i><0.05 versus PMD strain).</p

Western blot analysis of ALDH9948 and ALDH14080.

<p>(A) Elevated protein of ALDH9948 and ALDH14080 in PMD-R strain. Fifty micrograms of protein from New Orleans (NO), PMD and PMD-R strains in four life stages; larva (L), pupae (P), adult male (M) and adult female (F) including purified recombinant His-tagged ALDH9948 and 14080 (25 ng each) were resolved by SDS-PAGE. Proteins were transferred to a nitrocellulose membrane and probed with anti-ALDH9948 and anti-ALDH14080. Peroxidase labelled anti-rabbit antibody was used as a secondary antibody. Proteins were visualised by enhancing the chemiluminescence using ECL Advanced Blotting Detection Kit (Amersham Biosciences). Equal protein loading was confirmed by staining the membrane with Ponceus S. (B) Determination of antibodies specificity by western blot. Fifty nanograms of non-fusion ALDH9948 and ALDH14080 (Lane1, 2 and 3, respectively) were resolved in SDS-PAGE. Western blotting was performed as described.</p

Sequenced fragments of the Est6 gene for Anopheles scanloni mosquitoes

Sequenced fragments of the Est6 gene for Anopheles scanloni mosquitoe

Sequenced fragments of the Hsp82 gene for Anopheles scanloni mosquitoes

Sequenced fragments of the Hsp82 gene for Anopheles scanloni mosquitoe

Sequenced fragments of the Est6 gene for Anopheles baimaii mosquitoes

Sequenced fragments of the Est6 gene for Anopheles baimaii mosquitoe