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For our analysis, we apply three independent tools: matrix decomposition by singular value decomposition (SVD), supervised learning by agglomerative (hierarchical) clustering with Ward's method, and unsupervised learning by a self-organizing map (SOM, a type of neural network).
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As an alternative to these molecular tools, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) has gained interest for high throughput identification of microorganisms and has been established to identify adults and larvae of Culicoides[ 32, 33].
Database search was performed with the Mascot v2.1 search tool (Matrix Science, London, UK) screening SwissProt (release 56.0).
Matches to the known Mig3-binding matrix (Badis et al. 2008) were scored in the 800 bp upstream region using RSA-tools matrix scan (http://rsat.ulb.ac.be/) (Turatsinze et al. 2008); enrichment was estimated with Fisher's exact test, comparing with all genes in the genome with upstream matrix matches.
BACs were inoculated from a frozen stock plate using a 384-well pin tool (Matrix, CA) into microtiter plates containing 70 μl LB media plus 12.5 μg/ml chloramphenicol per well and the plates were incubated overnight at 37°C.
Among the matrix-based tools, stabilized matrix method (SMM) [ 75] and NetMHC [ 76] stand out for their performance [ 77, 78] and have been continuously upgraded.
Each TF peak set was scanned using the RSAT tool matrix-scan with PWMs for the four TFs.
Peaks for each TF were scanned using the RSAT tool matrix-scan with the best position weight matrices (PWM) for each TF.
Over-represented words (oligos) and spaced word pairs (dyads) were assembled and converted to position-specific scoring matrices with the tool matrix-from-patterns (RSAT).
Further, the tool matrix-scan from the RSAT toolbox determines putative TFBS in the promoter regions of target genes, which might be involved in transcriptional regulation [ 16].
The TRANSFAC formatted PSSMs were then employed to search for putative sites using the tool matrix-scan (http://www.rsat.eu/) [ 115, 116] with the determined optimal P-values set as the cut-offs.
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