Selection and validation of reference genes for gene expression studies in Klebsiella pneumoniae using Reverse Transcription Quantitative real-time PCR.

摘要: For reliable results, Reverse Transcription Quantitative real-time Polymerase Chain Reaction (RT-qPCR) analyses depend on stably expressed reference genes for data normalization purposes. Klebsiella pneumoniae is an opportunistic Gram-negative bacterium that has become a serious threat worldwide. Unfortunately, there no consensus ideal gene RT-qPCR K. pneumoniae. In this study, the expression profile of eleven candidate was assessed in cells submitted to various experimental conditions, and stability these evaluated using statistical algorithms BestKeeper, NormFinder, geNorm, Delta CT RefFinder. The ranked recA, rho, proC rpoD as most suitable accurate reliability proposed validated by normalizing relative iron-regulated iron-replete iron-limited conditions. This work emphasizes stable any potential must be each physiological condition or treatment under study.