Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
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微阵列引物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的适应
DOI：
10.1016/j.mex.2019.04.006
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发表时间：
2019
期刊：
影响因子：
1.9
通讯作者：
Bezbaruah, Achintya N.
中科院分区：
文献类型：
--
作者：
Fortuna, Ann-Marie;Sinha, Sanjivni;Das, Tonoy K.;Bezbaruah, Achintya N.
采用改进的PCR和培养方法对荧光假单胞菌细胞暴露于纳米零价铁（NZVI）后的荧光假单胞菌进行分析，并采用改进的原子吸收光谱（AAS）分析方法对细菌生长培养基中的铁（Fe）进行测定。我们采用了用于定量pvd和细菌铁蛋白相关铁氧还蛋白基因的基因表达的微阵列引物组用于实时定量逆转录（qRT-PCR）分析。pvd是调节铁载体绿脓菌荧光素合成的一组基因之一，也使用Chrome azrul S（CAS）平板测量。本方案提供了一种详细的qRT-PCR方法，用于定量参与Fe inP获得和利用的基因。荧光细胞暴露于NZVI。qRT-PCR结果通过2种基于培养的方法、生长曲线和Chrome azurol S（CAS）平板独立确证。·用改进的原子吸收法测定胰蛋白酶大豆肉汤（TSB）中的铁，其中钠（Na）会干扰铁的测定。
Modified 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.
DOI：
10.1007/978-94-007-6088-2
发表时间：
2013
期刊：
Iron Uptake in Bacteria with Emphasis on E. coli and Pseudomonas
影响因子：
--
作者：
R. Chakraborty
通讯作者：
R. Chakraborty
荧光假单胞菌从纳米铁吸收铁期间铁转运和稳态基因的表达减少
影响因子：
4.9
作者：
Sinha, Sanjivni;Das, Tonoy K.;Bezbaruah, Achintya N.;Fortuna, Ann-Marie
通讯作者：
Fortuna, Ann-Marie
影响因子：
2.9
作者：
SCHWYN, B;NEILANDS, JB
通讯作者：
NEILANDS, JB