sRNA phrs調(diào)控銅綠假單胞菌生物膜形成的功能研究
發(fā)布時(shí)間：2018-11-19 08:18
【摘要】：目的構(gòu)建銅綠假單胞菌(PA)小RNAphrs基因缺失株與回復(fù)株,觀察phrs在銅綠假單胞菌生長(zhǎng)和生物膜形成中的作用。方法應(yīng)用λ-Red重組系統(tǒng)構(gòu)建phrs缺失株P(guān)A27853Δphrs、回復(fù)株P(guān)A27853Δphrs-comp和對(duì)照株P(guān)A27853Δphrs-control,繪制細(xì)菌生長(zhǎng)曲線,觀察phrs對(duì)細(xì)菌生長(zhǎng)的影響;熒光染色法定量分析細(xì)菌生物膜形成能力變化。結(jié)果成功獲得phrs基因缺失株和回復(fù)株,RT-PCR結(jié)果顯示,phrs基因在缺失突變株中不表達(dá),在回復(fù)株中重新表達(dá);phrs不影響細(xì)菌的生長(zhǎng)速度,基因敲除后細(xì)菌生物膜形成能力明顯下降,而回復(fù)株則能形成明顯生物膜。結(jié)論利用λ-Red重組技術(shù)成功構(gòu)建出PA25783Δphrs,phrs對(duì)細(xì)菌生物膜的形成具有重要的調(diào)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.
【作者單位】： 中山大學(xué)孫逸仙紀(jì)念醫(yī)院醫(yī)院感染管理辦公室;中山大學(xué)孫逸仙紀(jì)念醫(yī)院檢驗(yàn)科;中山大學(xué)孫逸仙紀(jì)念醫(yī)院ICU科;
【基金】：廣東省科技計(jì)劃基金資助項(xiàng)目(2014A020212711)
【分類號(hào)】：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.
【作者單位】： 中山大學(xué)孫逸仙紀(jì)念醫(yī)院醫(yī)院感染管理辦公室;中山大學(xué)孫逸仙紀(jì)念醫(yī)院檢驗(yàn)科;中山大學(xué)孫逸仙紀(jì)念醫(yī)院ICU科;
【基金】：廣東省科技計(jì)劃基金資助項(xiàng)目(2014A020212711)
【分類號(hào)】：R378.991
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