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Open Access Research article

Plasmodium infection alters Anopheles gambiae detoxification gene expression

Rute C Félix1, Pie Müller234, Vera Ribeiro5, Hilary Ranson2 and Henrique Silveira1*

Author Affiliations

1 Centro de Malária e Outras Doenças Tropicais, UEI Malária, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Rua da Junqueira, 96, 1349-008 Lisbon, Portugal

2 Vector Group, Liverpool School of Tropical Medicine, Liverpool, L3 5QA, UK

3 Vector Control Unit, Medical Department, Swiss Tropical and Public Health Institute, CH-4002 Basel, Switzerland

4 University of Basel, CH-4003 Basel, Switzerland

5 Centro de Biomedicina Molecular e Estrutural (CBME), Instituto de Biotecnologia e Bioengenharia (IBB-LA), Universidade do Algarve, Faro, Portugal

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BMC Genomics 2010, 11:312  doi:10.1186/1471-2164-11-312

Published: 19 May 2010

Additional files

Additional file 1:

Table S1. Infection rate and oocyst load of A. gambiae infected with P. berghei used for the microarray experiments.

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Additional file 2:

Table S2. List of all the genes differentially expressed (p < 0.001) represented on the Detox chip including fold change in expression and p-values.

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Additional file 3:

Figure S1. Validation of the DNA microarray analysis using quantitative RT-PCR. The mean expression values for midgut genes (A) and fat body genes (B) obtained by microarray analysis were plotted against the corresponding mean expression values obtained with quantitative RT-PCR. A high level of consistency between the two datasets was demonstrated by the Pearson correlation coefficient (P = 0.884) for midgut and (P = 0.85) for fat body and best-fit linear-regression analysis (R2 = 0.7814) for midgut and (R2 = 0.7228) for fat body.

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Additional file 4:

Figure S2. Design of the microarray experiments. The experiments for midgut and fat body tissues followed the same layout. The boxes of the graphs represent RNA extracted from pools of 40 individuals and the arrows the microarrays to which labelled target RNA was co-hybridized. The tails of the arrows represent the samples that were labelled with a green (Cy3) and the heads those samples that were labelled with a red (Cy5) fluorescent dye. For the design matrix in limma, the samples from uninfected tissues collected 1 day post infection were set as the reference pool (shaded boxes). After fitting linear models the contrasts shown below the diagram were constructed for hypothesis testing of specific comparisons between RNA pools. For each of the three biological blocks (replicates 1 to 3) and factor combination a separate coefficient was included in the design matrix. The contrasts were extracted by taking the average of the three comparisons.

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Additional file 5:

Table S3. Sequences of oligonucleotide primers used in quantitative RT-PCR validation experiments.

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