摘要
目的观察新型mucA突变的粘液型菌株PA17和野生型菌株PAO1生物膜(biofilm,BF)形成的动态过程,并比较2株菌生物膜形成过程的差异。方法 SYTO9/PI荧光探针标记PAO1和PA17,体外建立1d,3d,5d,9d时间点PAO1及PA17的BF模型,激光共聚焦显微镜(CLSM)观察两株菌BF动态形成的过程。结果通过SYTO9/PI双染可以动态观察PAO1菌株和PA17菌株的BF形成过程;PAO1菌株和PA17菌株的BF形成过程有差异:PAO1菌株1d时已形成微菌落,3d时形成覆盖整个玻片的生物膜结构,而PA17菌株1d时仅有散在的不可逆粘附细菌,3d时才形成微菌落,5d时形成生物膜结构;随着时间的推移,2株菌生物膜形成的厚度均逐渐增加,且死菌的比例也逐渐增加。结论 PAO1和PA17BF的动态形成过程存在差异,而PA17与PAO1生物膜形成存在的差异可能与其mucA基因突变造成大量藻酸盐的产生有关。
Objective To observe the sequential biofilm development and compare the features of two strains of Pseudomonas aeruginosa： mucus-type PA17 with the new mucA gene mutation and non-mucoid PAO1 with wild-type mucA gene. Methods In vitro biofilm models of PAO1 and PA17 tagged with SYTO9/P1 were established on glass slices,and the biofilm development was monitored at different time points（1d,3d,5d and 9d）.The fluorescence images of different layers in the biofilm models were obtained by confocal laser scanning microscopy（CLSM） based on fluorophores from PAO1 and PA17. Results The PAO1 and PA17 biofilm development was investigated successfully by CLSM after the genetical tagging with SYTO9/P1.There were differences between PAO1 and PA17 in the development of the biofilm.The thickness of the biofilm was increased significantly during biofilm growth,and the ratio of dead bacteria was increased gradually in each layer as well. Conclusion The sequential development of biofilm formation is different between PAO1 and PA17,which may be associated with the mutation of the mucA gene in PA17.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2011年第5期448-451,共4页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家自然科学基金资助(3090127)
新教师基金资助(200804871069)
