新型mucA基因突变的铜绿假单胞菌生物被膜的形成和藻酸盐相关基因表达的研究  

The biofilm formation and the alginate biosynthetic genes expression of a Pseudomonas aeruginosa strain with a new type mucA gene mutation

作　　者：田德英[1] 倪明[1] 余冰[2] 雷洪波[1] 朱旭慧[3] 宋佩辉[1] 

机构地区：[1]华中科技大学同济医学院同济医院感染科,武汉430030 [2]华中科技大学同济医学院免疫学系 [3]华中科技大学同济医学院同济医院检验科

出　　处：《中华微生物学和免疫学杂志》2006年第1期14-17,共4页Chinese Journal of Microbiology and Immunology

基　　金：湖北省医药卫生科研基金资助(JX1B041)

摘　　要：目的 确定黏液型铜绿假单胞菌PA17的mum基因突变位点，研究藻酸盐合成相关基因在其生物被膜形成过程中的表达，并观察PAl7生物被膜形成过程和形态。方法 PCR方法扩增铜绿假单胞菌PA17的mueA基因全长并测序；改良平板培养法建立PA17的生物被膜模型，半定量RT-PCR测定生物被膜形成24h、3d．6d时藻酸盐合成相关基因,algD、algU和algR的表达，并进行统计学分析；扫描电镜观察不同时间点的生物被膜形态。结果 PA17的mucA基因第166～333位核苷酸片段缺失，第342位A→G；其藻酸盐相关基因algD和algU均在生物被膜形成过程的第6天表达水平最高，algR在24h表达最高，单因素方差分析显示，上述基因在生物被膜形成过程不同时间点表达的差异有统计学意义；PA17于第6天形成成熟生物被膜，形态为薄膜状。结论 PA17是一株含新型mucA突变基因的黏液型铜绿假单胞菌，其藻酸盐相关基因在生物被膜形成的不同时间点的表达差异具有统计学意义，其生物被膜形态为薄膜状。Objective To identify the mutation point of Pseudomonas aeruginosa strain PA17 and to investigate the alginate biosynthetic genes expression during biofilm formation, Methods The mucA gene of PA17 was amplified and the products were sequenced. A modified plate culture method was used to establish the biofilm model of PA17. Semi-quantitative RT-PCR was used to determine the expresaion level of algD, algU and algR during the biofilm formation. One-Way ANOVA was used to identify the significance of the difference. Scanning electron microscope was used to observe the biofilm structure. Results There was a 166-333 deletion mutation and 342 A→G in mucA gene of PA17. The expression level of algD and algU was maximal at the 6 d time point during biofilm formation but the expression level of algR was maximal at 24 h time point. The expression difference of algD, algU and algR in different time point of biofilm formation had statistic significance. PA17 biofilm was mature after 6 d and the biofilm structure is a pellicle-shaped one. Conclusion PA17 is a mueoid Pseudomonas aeruginosa strain with a new type of mutation of mucA gene, and the expression difference of alginate biosynthetic genes of this strain in different time points of biofilm formation had statistic significance, the biofilm structure of PA17 is a pellicle-shaped one.

关 键 词：铜绿假单胞菌 mucA基因 生物被膜 藻酸盐 

分 类 号：R378[医药卫生—病原生物学]