gingivalis W83 genome. Before our study PF-4708671 all probes were analyzed for their unique- and perfect matching with the genome, as downloaded from the NCBI, using BLAST. Twenty-nine of the 1907 probes of the microarray gave non-specific hits, mostly related to transposases (Table 2). These probes were excluded from further analyses together with four probes that were not in use anymore annotated “”GSK1838705A purchase obsolete”" by the manufacturer, so that 1874 probes remained. The comparison of each test strain to W83 using this array gives insights into described virulence associated genes. A limitation
of the method, however, is that genes from the variable gene pool from other strains will not be detected. Table 2 Probes excluded from analysis due to redundancy GeneID Annotated function PG2152 CCI-779 datasheet DNA-binding protein, histone-like family PG0261 ISPg3, transposase PG0943 ISPg5, transposase Orf2 PG1420 ISPg5, transposase Orf2 PG1444 hypothetical protein PG1261 ISPg4, transposase PG1276 DNA-binding
protein, histone-like family PG1670 hypothetical protein PG1451 conserved hypothetical protein PG2128 ISPg5, transposase Orf2 PG1449 conserved hypothetical protein PG1453 Integrase PG1267 hypothetical protein PG1350 ISPg2, transposase PG0827 MATE efflux family protein PG1669 hypothetical protein PG1448 ISPg1, transposase PG1709 ISPg5, transposase Orf1 PG1454 Integrase PG1332 NAD(P) transhydrogenase, beta subunit PG1452 lipoprotein, putative PG1384 ISPg1, transposase, authentic frameshift PG1244 ISPg1, transposase PG1447 transcriptional regulator, AraC family PG1450 conserved hypothetical protein PG1445
rteC protein, truncation PG1671 hypothetical protein PG0487 ISPg4, transposase PG0760 ISPg1, transposase, authentic frameshift Data were normalized and technical and biological replicates were collapsed as described in the Materials and Methods. Detailed analysis G protein-coupled receptor kinase of the probe intensities indicated that 22 probes gave systematically low intensity values for strain W83 as well as for all the other strains. The intensity levels were at the same low levels as the intensity levels of the negative control probes (Figure 1). These probes were labeled as “”dead probes”" and excluded from the results (Table 3). Our data do not explain why dead probes have occurred in our experiments, but the consistent low signal for these probes suggests that the sequencing information used for designing these probes was imperfect. Figure 1 Hybridization signals of P. gingivalis strains – dead probes. A. The total intensity distribution of probe signals of W83 DNA hybridized to the W83 array. The density peak around 7.5 contains the negative controls (empty spots and A. thaliana probes). The peak around 12 should contain all present genes in strain W83. B Probe signal intensities of each P. gingivalis test strain are represented in light blue dots; medium blue dots, slightly below that, symbolize A. thaliana negative control genes.