sRNA phrs调控铜绿假单胞菌生物膜形成的功能研究  

Function of small RNA phrs on modulating biofilm formation in Pseudomonas aeruginosa

作　　者：徐凤琴[1] 黄松音[2] 莫红平[3] 曾俏君 孙婧[1] 周春霞[1] 李红玉[2] 

机构地区：[1]中山大学孙逸仙纪念医院医院感染管理办公室,广东广州510120 [2]中山大学孙逸仙纪念医院检验科,广东广州510120 [3]中山大学孙逸仙纪念医院ICU科,广东广州510120

出　　处：《中华医院感染学杂志》2017年第10期2169-2172,共4页Chinese Journal of Nosocomiology

基　　金：广东省科技计划基金资助项目(2014A020212711)

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。OBJECTIVE To construct small RNA phrs gene deletion and complementation strains from Pseudomonas aeruginosa and observe the effect of the phrs on the bacterial growth,and biofilm formation.METHODS The λ-Red recombination system was used to generate the phrs deletion strain （PA27853△phrs）,complementary strain （PA27853△phrs-comp）,and control strain （PA27853△phrs-control） of P.aeruginosa.The growth curves were drawn to assess the effect of phrs on the bacterial growth.Fluorescent staining assay was carried out for quantitative analysis of the change of ability of biofilm formation.RESULTS The phrs deletion and complementation strains were constructed successfully.The RT-PCR showed that the phrs gene was re-expressed in complementary strain but was not expressed in deletion strain.The phrs gene did not have effect on the speed of bacterial growth,the ability of biofilm formation was remarkably reduced after the gene deletion,however,the biofilms were remarkably formed in the complementation strains.CONCLUSION PA27853△phrs strain is successfully constructed by λ-Red recombination technology.The phrs gene plays an important role in regulating the biofilm formation of P.aeruginosa.

关 键 词：铜绿假单胞菌 生物膜 小RNA 

分 类 号：R378.991[医药卫生—病原生物学]