Open Access Research article

Transcriptomic and functional analysis of the Anopheles gambiae salivary gland in relation to blood feeding

Suchismita Das1, Andrea Radtke1, Young-Jun Choi2, Antonio M Mendes13, Jesus G Valenzuela4 and George Dimopoulos1*

Author Affiliations

1 W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, 615 N Wolfe Street, Baltimore, MD 21205-2179, USA

2 Department of Pathobiological Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA

3 Imperial College London, Division of Cell and Molecular Biology, Faculty of Natural Sciences, South Kensington Campus, London, UK

4 Laboratory of Malaria and Vector Research, NIAID, National Institutes of Health, Rockville, Maryland 20852, USA

For all author emails, please log on.

BMC Genomics 2010, 11:566  doi:10.1186/1471-2164-11-566

Published: 14 October 2010

Additional files

Additional file 1:

List of genes expressed in the female salivary gland of A. gambiae, along with their previous (ENSANGT) and recent (AGAP-R) transcript IDs. Note that the recent AGAP-R IDs for some transcripts are missing in VectorBase and have therefore been left blank in column B. The fluorescent values at 635 nm (Cy-5 dye) and 532 nm (Cy-3 dye) for each gene are shown in column E and F respectively. The excel file has three sheets: Sheet 1 lists the genes that are highly expressed, with fluorescent values (at 635 nm) from 5,000 to the maximum threshold of 65,000, Sheet 2 lists the moderately expressed genes with fluorescent values (at 635 nm) between 1,000 to 4,999 and Sheet 3 lists the weakly expressed genes with fluorescent values (at 635 nm) between 100 to 999. The gene list is further sorted into different functional groups, according to the predicted function of the genes [CS: cytoskeletal and structural; CIR: circadian; CSR: chemosensory; PROT/DIG: proteolytic digestion; IMM: immunity; MET/ENZ: metabolism and enzymes; RED/STR: redox/stress; R/T/T: replication/transcription and translation; TRP: transport]. The list of genes in each group is further sorted into descending order of the fluorescent value at 635 nm (column E).

Format: XLS Size: 846KB Download file

This file can be viewed with: Microsoft Excel Viewer

Open Data

Additional file 2:

List of genes that are regulated upon blood-feeding in the female A. gambiae salivary gland, along with their ENSANGT and more recent AGAP-R transcript IDs. Transcripts lacking AGAP-R IDs are only indicated with the ENSANT transcript ID. Genes are only listed if they display significant differential regulation between fed and unfed mosquitoes, at a log2 fold ratio above or below the ± 0.75 threshold. Predicted functional groups classification for genes are also presented [CS: cytoskeletal and structural; CIR: circadian; CSR: chemosensory; PROT/DIG: proteolytic digestion; IMM: immunity; MET/ENZ: metabolism and enzymes; RED/STR: redox/stress; R/T/T: replication/transcription and translation; TRP: transport].

Format: XLS Size: 31KB Download file

This file can be viewed with: Microsoft Excel Viewer

Open Data

Additional file 3:

Gene silencing efficiency of the 10 selected genes. The gene transcript abundance in GFP dsRNA treated mosquitoes was set to 1.0, and the corresponding percentage silencing was determined by qRT-PCR. The AgS7 gene was used for normalization of cDNA templates. The standard error values are shown.

Format: DOC Size: 36KB Download file

This file can be viewed with: Microsoft Word Viewer

Open Data

Additional file 4:

The blood-feeding propensity of mosquitoes in gene silenced mosquitoes. Ten salivary gland expressed genes were individually silenced in mosquitoes, and the mosquitoes were allowed to feed on blood, 4 days after dsRNA injections. The percentages of mosquitoes that feed ("Fed") were scored and are shown along with their standard error values. The p-value from statistical analysis of Mann Whitney Test is also shown.

Format: DOC Size: 38KB Download file

This file can be viewed with: Microsoft Word Viewer

Open Data

Additional file 5:

Primers used for synthesis of dsRNAs for RNAi gene silencing assays. The first 20 bases in bold correspond to the T7 polymerase promoter site. The transcript ID numbers (AGAP-RA from VectorBase) are also shown for each gene. The list also includes the sequences of GFP primers used for generating dsRNA and the A. gambiae S7 primer (used for standardization of cDNA templates).

Format: DOC Size: 36KB Download file

This file can be viewed with: Microsoft Word Viewer

Open Data

Additional file 6:

Primers used for verification of RNAi silencing. The sense primers were newly designed (VERI Sense), and the antisense primers used were the same as those used for making the corresponding dsRNAs. The transcript ID numbers (AGAP-RA from VectorBase) are also shown for each gene.

Format: DOC Size: 34KB Download file

This file can be viewed with: Microsoft Word Viewer

Open Data