Orthologous genes were identified as best hits using blastp analy

Orthologous genes were identified as best hits using blastp analysis (blastall v2.2.22) [71, 72] against local databases. Cut-offs of 50% identity over at least 80% of the sequence length and an expected value (e-value) of 1e-10 were applied. Orthology was confirmed by reciprocating the blastp analysis. Since the A-rich motif is short and degenerate it is expected that occurrences of the A-rich motif that are unrelated to Crc binding will be detected in this analysis, giving rise to false positive hits. In order to estimate

the rate of false positive hits in our analysis we searched for the A-rich motif in the AZD2171 manufacturer reverse Selleck LY3023414 orientation of the upstream regions of orthologous loci [73]. Since the A-rich motif in the reverse orientation is unrelated to Crc binding it is reasoned that this estimates the rate of occurrence of the A-rich motif in the sequence fragments tested. Predictably it was found that the use of more strains per species resulted in lower estimated rates of false positives (P. aeruginosa – 4 strains, 18% estimated false positives; P. fluorescens – 3 strains, Angiogenesis inhibitor 32% estimated false positives; P. putida – 3 strains, 26% estimated false positives; P. syringae – 2 strains, 41% estimated

false positives). Thus, it is estimated, based on the weighted mean false discovery rate, that approximately 73% of the Crc candidates in additional file 1 are genuine targets for Crc binding. Functional information about the translated protein sequences was obtained from the sequence headers Teicoplanin and by performing Blast2GO analysis [74]. Acknowledgements This research was supported in part by grants awarded by the Science Foundation of Ireland (grants 04/BR/B0597, 07/IN.1/B948, 08/RFP/GEN1295, 08/RFP/GEN1319 and 09/RFP/BMT2350), the Department of Agriculture, Fisheries and Food (RSF grants 06-321 and 06-377; FIRM grants 06RDC459, 06RDC506 and 08RDC629),

the European Commission (grant FP6#O36314 and Marie Currie TOK:TRAMWAYS), Irish Research Council for Science Engineering and Technology (grant 05/EDIV/FP107/INTERPAM), the Marine Institute (Beaufort award C&CRA2007/082), the Health Research Board (grants RP/2006/271 and RP/2007/290). P.B. is supported by a STRIVE Doctoral Scholarship from the Environmental Protection Agency, Ireland and the Department of Environment, Heritage and Local Government provided by the Irish Government under the National Development Plan 2007-2013 (EPA-2006-S-21). We thank Pat Higgins for ongoing techncial support and members of our groups for useful discussions. Electronic supplementary material Additional file 1: Crc candidates identified in every Pseudomonas spp. List of every locus bearing a Crc motif in P. aeruginosa, P. fluorescens, P. putida and P. syringae species. The numbers under strain names on the left indicate the locus id, according to Genbank annotation, of the locus with the A-rich motif in the upstream region.

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