sRNA phrs调控铜绿假单胞菌生物膜形成的功能研究
发布时间：2018-11-19 08:18
【摘要】：目的构建铜绿假单胞菌(PA)小RNAphrs基因缺失株与回复株,观察phrs在铜绿假单胞菌生长和生物膜形成中的作用。方法应用λ-Red重组系统构建phrs缺失株PA27853Δphrs、回复株PA27853Δphrs-comp和对照株PA27853Δphrs-control,绘制细菌生长曲线,观察phrs对细菌生长的影响;荧光染色法定量分析细菌生物膜形成能力变化。结果成功获得phrs基因缺失株和回复株,RT-PCR结果显示,phrs基因在缺失突变株中不表达,在回复株中重新表达;phrs不影响细菌的生长速度,基因敲除后细菌生物膜形成能力明显下降,而回复株则能形成明显生物膜。结论利用λ-Red重组技术成功构建出PA25783Δphrs,phrs对细菌生物膜的形成具有重要的调控作用。
[Abstract]:Objective to construct the small RNAphrs gene deletion and recovery strain of Pseudomonas aeruginosa (PA) and to observe the role of phrs in the growth and biofilm formation of Pseudomonas aeruginosa. Methods A 位-Red recombinant system was used to construct phrs deletion strain PA27853 螖 phrs, PA27853 螖 phrs-comp and control strain PA27853 螖 phrs-control, to draw the bacterial growth curve, to observe the effect of phrs on bacterial growth, and to quantitatively analyze the change of bacterial biofilm formation ability by fluorescence staining. Results phrs gene deletion and response strain were successfully obtained. RT-PCR results showed that phrs gene was not expressed in the deletion mutant, but was reexpressed in the response strain. Phrs did not affect the growth rate of bacteria, but the ability of biofilm formation was significantly decreased after gene knockout, while the biofilm was formed in reverse-strain. Conclusion PA25783 螖 phrs,phrs was successfully constructed by 位-Red recombination technique to regulate the formation of bacterial biofilm.
【作者单位】： 中山大学孙逸仙纪念医院医院感染管理办公室;中山大学孙逸仙纪念医院检验科;中山大学孙逸仙纪念医院ICU科;
【基金】：广东省科技计划基金资助项目(2014A020212711)
【分类号】：R378.991
[Abstract]:Objective to construct the small RNAphrs gene deletion and recovery strain of Pseudomonas aeruginosa (PA) and to observe the role of phrs in the growth and biofilm formation of Pseudomonas aeruginosa. Methods A 位-Red recombinant system was used to construct phrs deletion strain PA27853 螖 phrs, PA27853 螖 phrs-comp and control strain PA27853 螖 phrs-control, to draw the bacterial growth curve, to observe the effect of phrs on bacterial growth, and to quantitatively analyze the change of bacterial biofilm formation ability by fluorescence staining. Results phrs gene deletion and response strain were successfully obtained. RT-PCR results showed that phrs gene was not expressed in the deletion mutant, but was reexpressed in the response strain. Phrs did not affect the growth rate of bacteria, but the ability of biofilm formation was significantly decreased after gene knockout, while the biofilm was formed in reverse-strain. Conclusion PA25783 螖 phrs,phrs was successfully constructed by 位-Red recombination technique to regulate the formation of bacterial biofilm.
【作者单位】： 中山大学孙逸仙纪念医院医院感染管理办公室;中山大学孙逸仙纪念医院检验科;中山大学孙逸仙纪念医院ICU科;
【基金】：广东省科技计划基金资助项目(2014A020212711)
【分类号】：R378.991
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