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Background  Reverse transcription quantitative PCR has become a powerful technique to monitor mRNA transcription in response to different environmental conditions in many bacterial species. However, correct evaluation of data requires accurate and reliable use of reference genes whose transcription does not change during the course of the experiment. In the present study exposure to different growth conditions was used to validate the transcription stability of eight reference gene candidates in three strains from two subspecies of  Francisella noatunensis , a pathogen causing disease in both warm and cold water fish species.    Results  Relative transcription levels for genes encoding DNA gyrase ( gyrA ), RNA polymerase beta subunit ( rpoB ), DNA polymerase I ( polA ), cell division protein ( ftsZ ), outer membrane protein ( fopA ), riboflavin biosynthesis protein ( ribC ), 16S ribosomal RNA ( 16S rRNA ) and DNA helicases ( uvrD ) were quantified under exponential, stationary and iron-restricted growth conditions. The suitability of selected reference genes for reliable interpretation of gene expression data was tested using the virulence-associated intracellular growth locus subunit C ( iglC ) gene.    Conclusion  Although the transcription stability of the reference genes was slightly different in the three strains studied,  fopA ,  ftsZ  and  polA  proved to be the most stable and suitable for normalization of gene transcription in  Francisella noatunensis  ssp.

bmc research notes

Evaluation of reference genes for reverse transcription quantitative PCR analyses of fish-pathogenic Francisella strains exposed to different growth conditions