Validation of array data by real-time PCRa

Gene identifierGeneQuantitative real-time RT-PCRb result (relative difference in expression change)Expression difference of the hemB mutant analyses compared to the wild type using microarraysc
hemB mutantComplemented mutantWild type
SA0150Capsular polysaccharide synthesis enzyme Cap5G29.91.11.0Sig. up-reg.
SA0232l-Lactate dehydrogenase21.11.01.2Sig. up-reg.
SA0742Fibrinogen-binding protein A494.612.61.0Sig. up-reg.
SA0922Phosphoribosylpyrophosphate amidotransferase PurF1.024.310.6Sig. down-reg.
SA1141Glycerol kinase40.83.61.0Sig. up-reg.
SA1142Aerobic glycerol-3-phosphate dehydrogenase128.01.02.4Sig. up-reg.
SA2206Immunoglobulin G-binding protein SBI1.04.38.3Sig. down-reg.
SA2424Hypothetical protein, similar to transcription regulator Crp/Fnr family protein163.11.01.3Sig. up-reg.
SA2426Arginine/ornithine antiporter891.01.01.3Sig. up-reg.
SA2428Arginine deiminase989.02.11.0Sig. up-reg.
  • a Transcription rates of selected genes for the S. aureus hemB mutant in comparison to the wild type and the complemented mutant for pooled time points. The relative differences in expression change were calculated using Gene Expression Analysis for iCycler iQ Real-Time PCR Detection System v1.10 (Bio-Rad). The lowest expression was set to 1.0. gyrA (SA0006) was used as a housekeeping gene for relative quantification.

  • b RT-PCR, reverse transcription-PCR.

  • c See Table 2. Abbreviations: sig. up-reg., significantly up-regulated; sig down-reg., significantly down-regulated.