Wang Yao, Ding LiSha, Hu YangBo, Zhang Yong, Yang BaoYu, Chen ShiYun
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Sci China C Life Sci. 2007 Dec;50(6):814-21. doi: 10.1007/s11427-007-0101-6.
The flagella master regulatory gene flhDC of Yersinia pseudotuberculosis serotype III (YPIII) was mutated by deleting the middle region and replaced by a tetracycline resistant gene, and the subsequent mutant strain named YPIIIDeltaflhDC was obtained. Swimming assay showed that the swimming motility of the mutant strain was completely abolished. The promoter region of the flagella second-class regulatory gene fliA was fused with the lux box, and was conjugated with the mutant and the parent strains respectively for the first cross. LUCY assay result demonstrated that flhDC regulated the expression of fliA in YPIII as reported in E. coli. Biofilm formation of the mutant strain on abiotic and biotic surfaces was observed and quantified. The results showed that mutation of flhDC decreased biofilm formation on both abiotic and biotic surfaces, and abated the infection on Caenorhabdtis elegans. Our results suggest that mutation of the flagella master regulatory gene flhDC not only abolished the swimming motility, but also affected biofilm formation of YPIII on different surfaces. The new function of flhDC identified in this study provides a novel viewpoint for the control of bacterial biofilm formation.
通过缺失中间区域对假结核耶尔森菌III型(YPIII)的鞭毛主调控基因flhDC进行突变,并用四环素抗性基因进行替换,从而获得了后续的突变菌株,命名为YPIIIDeltaflhDC。游动试验表明,该突变菌株的游动能力完全丧失。将鞭毛二级调控基因fliA的启动子区域与lux盒融合,并分别与突变菌株和亲本菌株进行第一次杂交。LUCY试验结果表明,flhDC如在大肠杆菌中报道的那样调控YPIII中fliA的表达。观察并定量了突变菌株在非生物和生物表面上的生物膜形成情况。结果表明,flhDC的突变降低了在非生物和生物表面上的生物膜形成,并减轻了对秀丽隐杆线虫的感染。我们的结果表明,鞭毛主调控基因flhDC的突变不仅消除了游动能力,还影响了YPIII在不同表面上的生物膜形成。本研究中鉴定出的flhDC的新功能为控制细菌生物膜形成提供了新的视角。