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Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays

Pseudomonas aeruginosa is an opportunistic human pathogen notorious for its remarkable capacity of multi-drug resistance, and has become one of the most important model bacteria in clinical bacteriology research. Quantitative real-time PCR is a reliable method widely used in gene expression analysis...

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Autores principales: Meng, Lingning, Cao, Xiaoli, Li, Chuchu, Li, Jia, Xie, Hui, Shi, Jiping, Han, Mei, Shen, Han, Liu, Chang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10040570/
https://www.ncbi.nlm.nih.gov/pubmed/36992935
http://dx.doi.org/10.3389/fmicb.2023.1140515
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author Meng, Lingning
Cao, Xiaoli
Li, Chuchu
Li, Jia
Xie, Hui
Shi, Jiping
Han, Mei
Shen, Han
Liu, Chang
author_facet Meng, Lingning
Cao, Xiaoli
Li, Chuchu
Li, Jia
Xie, Hui
Shi, Jiping
Han, Mei
Shen, Han
Liu, Chang
author_sort Meng, Lingning
collection PubMed
description Pseudomonas aeruginosa is an opportunistic human pathogen notorious for its remarkable capacity of multi-drug resistance, and has become one of the most important model bacteria in clinical bacteriology research. Quantitative real-time PCR is a reliable method widely used in gene expression analysis, for which the selection of a set of appropriate housekeeping genes is a key prerequisite for the accuracy of the results. However, it is easy to overlook that the expression level of housekeeping gene may vary in different conditions, especially in the condition of molecular microbiology assays, where tested strains are generally cultured under the pre-set antibiotic selection pressures, and how this affects the stability of commonly used housekeeping genes remains unclear. In this study, the expression stability of ten classic housekeeping genes (algD, gyrA, anr, nadB, recA, fabD, proC, ampC, rpoS, and rpsL) under the pressure of eight laboratory commonly used antibiotics (kanamycin, gentamycin, tetracycline, chloramphenicol, hygromycin B, apramycin, tellurite, and zeocin) were tested. Results showed that the stability of housekeeping gene expression was indeed affected by the types of antibiotics added, and of course the best reference gene set varied for different antibiotics. This study provides a comprehensive summary of the effects of laboratory antibiotics on the stability of housekeeping genes in P. aeruginosa, highlighting the necessity to select housekeeping genes according to the type of antibiotics used in the initial stage of experiment.
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spelling pubmed-100405702023-03-28 Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays Meng, Lingning Cao, Xiaoli Li, Chuchu Li, Jia Xie, Hui Shi, Jiping Han, Mei Shen, Han Liu, Chang Front Microbiol Microbiology Pseudomonas aeruginosa is an opportunistic human pathogen notorious for its remarkable capacity of multi-drug resistance, and has become one of the most important model bacteria in clinical bacteriology research. Quantitative real-time PCR is a reliable method widely used in gene expression analysis, for which the selection of a set of appropriate housekeeping genes is a key prerequisite for the accuracy of the results. However, it is easy to overlook that the expression level of housekeeping gene may vary in different conditions, especially in the condition of molecular microbiology assays, where tested strains are generally cultured under the pre-set antibiotic selection pressures, and how this affects the stability of commonly used housekeeping genes remains unclear. In this study, the expression stability of ten classic housekeeping genes (algD, gyrA, anr, nadB, recA, fabD, proC, ampC, rpoS, and rpsL) under the pressure of eight laboratory commonly used antibiotics (kanamycin, gentamycin, tetracycline, chloramphenicol, hygromycin B, apramycin, tellurite, and zeocin) were tested. Results showed that the stability of housekeeping gene expression was indeed affected by the types of antibiotics added, and of course the best reference gene set varied for different antibiotics. This study provides a comprehensive summary of the effects of laboratory antibiotics on the stability of housekeeping genes in P. aeruginosa, highlighting the necessity to select housekeeping genes according to the type of antibiotics used in the initial stage of experiment. Frontiers Media S.A. 2023-03-13 /pmc/articles/PMC10040570/ /pubmed/36992935 http://dx.doi.org/10.3389/fmicb.2023.1140515 Text en Copyright © 2023 Meng, Cao, Li, Li, Xie, Shi, Han, Shen and Liu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Meng, Lingning
Cao, Xiaoli
Li, Chuchu
Li, Jia
Xie, Hui
Shi, Jiping
Han, Mei
Shen, Han
Liu, Chang
Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays
title Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays
title_full Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays
title_fullStr Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays
title_full_unstemmed Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays
title_short Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays
title_sort housekeeping gene stability in pesudomonas aeruginosa pao1 under the pressure of commonly used antibiotics in molecular microbiology assays
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10040570/
https://www.ncbi.nlm.nih.gov/pubmed/36992935
http://dx.doi.org/10.3389/fmicb.2023.1140515
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