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

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 ada...

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Autores principales: Fortuna, Ann-Marie, Sinha, Sanjivni, Das, Tonoy K., Bezbaruah, Achintya N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6535460/
https://www.ncbi.nlm.nih.gov/pubmed/31193529
http://dx.doi.org/10.1016/j.mex.2019.04.006
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author Fortuna, Ann-Marie
Sinha, Sanjivni
Das, Tonoy K.
Bezbaruah, Achintya N.
author_facet Fortuna, Ann-Marie
Sinha, Sanjivni
Das, Tonoy K.
Bezbaruah, Achintya N.
author_sort Fortuna, Ann-Marie
collection PubMed
description 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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spelling pubmed-65354602019-05-30 Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron Fortuna, Ann-Marie Sinha, Sanjivni Das, Tonoy K. Bezbaruah, Achintya N. MethodsX Environmental Science 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. Elsevier 2019-04-30 /pmc/articles/PMC6535460/ /pubmed/31193529 http://dx.doi.org/10.1016/j.mex.2019.04.006 Text en © 2019 Published by Elsevier B.V. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Environmental Science
Fortuna, Ann-Marie
Sinha, Sanjivni
Das, Tonoy K.
Bezbaruah, Achintya N.
Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
title Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
title_full Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
title_fullStr Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
title_full_unstemmed Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
title_short Adaption of microarray primers for iron transport and homeostasis gene expression in Pseudomonas fluorescens exposed to nano iron
title_sort adaption of microarray primers for iron transport and homeostasis gene expression in pseudomonas fluorescens exposed to nano iron
topic Environmental Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6535460/
https://www.ncbi.nlm.nih.gov/pubmed/31193529
http://dx.doi.org/10.1016/j.mex.2019.04.006
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