Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron

针对暴露于纳米铁的荧光假单胞菌中铁转运和稳态基因表达,对微阵列引物进行改造

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Abstract

Modified protocols were adapted for PCR and culture based methods for the analysis of Pseudomonas fluorescens cells exposed to nanoscale zero-valent iron (NZVI) and iron (Fe) in bacterial growth nutrient media was determined by a modified atomic absorption spectrometric (AAS) analysis method. We adapted sets of microarray primers used to quantify gene expression of pvdS and a bacterioferritin-associated ferredoxin gene for use in real-time quantitative reverse transcription (qRT-PCR) analysis. pvdS is one of a cluster of genes regulating the synthesis of the siderophore pyoverdine that was also measured using chrome azrul S (CAS) plates. •The current protocol provides a detailed qRT-PCR method for quantifying genes involved in the acquisition and utilization of Fe in P. fluorescens cells exposed to NZVI.•The qRT-PCR results were independently corroborated with 2 culture based methods, growth curves and chrome azurol S (CAS) plate.•The modified AAS method was used to measure Fe in Tryptic Soy Broth (TSB) medium where sodium (Na) causes inference in iron measurement.

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