文章基本信息

标题：Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
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作者：Ann-Marie Fortuna ; Sanjivni Sinha ; Tonoy K. Das 等
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2019
卷号：6
页码：1181-1187
DOI：10.1016/j.mex.2019.04.006
语种：English
出版社：Elsevier
摘要：Graphical abstractDisplay OmittedAbstractModified protocols were adapted for PCR and culture based methods for the analysis ofPseudomonas fluorescenscells 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 ofpvdSand a bacterioferritin-associated ferredoxin gene for use in real-time quantitative reverse transcription (qRT-PCR) analysis.pvdSis 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 inP. fluorescenscells 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.
关键词：qRT-PCR;Azrul S (CAS);Atomic absorption;Pyoverdine;Ferredoxin