Results and discussion Comparative transcriptomics The clinical VISA isolate SA137/93A and the type strain of the Iberian clone in Germany (‘Northern German epidemic strain’) SA1450/94 CYT387 showed identical PFGE patterns [43] and MLST types (ST247). In order to further confirm that SA1450/94
is a suitable control strain, chromosomal DNA of SA137/93A was competitively hybridised to that of SA1450/94 and SA137/93G, respectively. The microarray results showed that all ORFs present in the VISA strain SA137/93A were also present in strain SA1450/94. In addition, the competitive hybridisation precisely reflected the deletion in the mutant SA137/93G [4]. Comparative Selleckchem INCB28060 transcriptomics of the hVISA isolate SA137/93A and SA1450/94 revealed that there were only 15 genes showing a higher expression level in the hVISA strain (2- to12-fold; see Additional file 1: gene expression data.pdf, Additional file 1: Table S1). The yycFGHI-operon [27] and three genes of the type 5/8 capsule biosynthesis gene cluster (capB, capC, capE),
which showed a 4- to 5-fold higher expression, were among the ten genes with a known function in Additional file 1: Table S1. The relatively low number of regulated genes may be due to the fact that the strain shows a heterogeneous phenotype, i.e., only a subpopulation of the culture displays high resistance to vancomycin. Similar results were obtained with the JH series of mutants; here JH1 to JH5 did not show any alterations in gene expression, although resistance had increased [44, 45], therefore, this observation was not surprising. Selleck Semaxanib As expected, the transcription profile of the VISA strain SA137/93G differed more strikingly from that of SA1450/94. A total of 124 genes showed at least a twofold change in strain SA137/93G (2- to 13.7-fold; see Additional file 1: gene expression data.pdf, Additional file 1: Table S2) compared to SA1450/94. 30.6% of these genes encoded hypothetical
proteins. Figure MEK inhibitor 1 shows the percentage of regulated genes in the different functional gene classes. Only one category of genes, “adaptation to atypical conditions”, which comprises genes encoding capsule biosynthesis enzymes, chaperones, heat and cold shock proteins and the clp protease, was overrepresented among the genes showing higher transcription levels. The 16 genes of the capsular biosynthesis gene cluster capABCDEFGHIJKLMNOP were – on average – six fold up-regulated. Additionally, a more than twofold increase in transcript amounts was found for the gene encoding AlsS, which is involved in formation of acetoine from pyruvate and influences the regulation of autolysis [46], the urease operon and two ORFs of the ica gene cluster. All of the above mentioned genes have also been found to be up-regulated under mildly acidic conditions [47].