Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays.
Housekeeping gene stability in Pesudomonas aeruginosa PAO1 under the pressure of commonly used antibiotics in molecular microbiology assays.
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DOI：
10.3389/fmicb.2023.1140515
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发表时间：
2023
影响因子：
5.2
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中科院分区：
文献类型：
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铜绿假单胞菌是一种机会致病菌，具有显著的多重耐药能力，已成为临床细菌学研究的重要模式菌之一。实时定量聚合酶链式反应是一种广泛应用于基因表达分析的可靠方法，选择一组合适的看家基因是保证结果准确的关键前提。然而，人们容易忽视的是，看家基因的表达水平在不同的条件下可能会有所不同，特别是在分子微生物学检测的条件下，测试菌株通常是在预先设定的抗生素选择压力下培养的，这如何影响常用管家基因的稳定性尚不清楚。本研究测试了10个经典看家基因(algD、gyra、anr、nadB、recA、faD、proc、ampC、rpos和rpsL)在8种实验室常用抗生素(卡那霉素、庆大霉素、四环素、氯霉素、潮霉素B、安普霉素、亚碲酸盐和玉米素)压力下的表达稳定性。结果表明，家政基因表达的稳定性确实受到添加的抗生素种类的影响，当然，不同抗生素的最佳参考基因集是不同的。本研究对实验室抗生素对铜绿假单胞菌看家基因稳定性的影响进行了全面的总结，强调了根据实验初期使用的抗生素类型选择看家基因的必要性。
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.