Wang Yao, Dai Yue, Zhang Yong, Hu YangBo, Yang BaoYu, Chen ShiYun
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Sci China C Life Sci. 2007 Jun;50(3):385-91. doi: 10.1007/s11427-007-0044-y.
The aiiA gene from Bacillus thuringiensis was cloned into the Pseudomonas/E. coli shuttle vector and transformed into Pseudomonas aeruginosa strain PAO1. Western blotting showed that the AiiA protein was expressed in PAO1. After induction by IPTG for 6 h and 18 h, expression of the aiiA gene in PAO1 completely degraded the quorum sensing autoinducers N-acylhomoserine lactones (AHLs): N-oxododecanoyl-L-homoserine lactone (OdDHL) and N-butyryl-L-homoserine lactone (BHL). The reduced amount of AHLs in PAO1 was also correlated with decreased expression and production of several virulence factors such as elastase and pyocyanin. AiiA expression also influenced bacterial swarming motility. Most importantly, our studies indicated that aiiA played significant roles in P. aeruginosa biofilm formation and dispersion, as observed by the differences of the biofilm formation on liquid and solid surfaces, and biofilm structures under a scanning electron microscope.
将来自苏云金芽孢杆菌的aiiA基因克隆到假单胞菌/大肠杆菌穿梭载体中,并转化到铜绿假单胞菌PAO1菌株中。蛋白质免疫印迹法显示AiiA蛋白在PAO1中表达。经异丙基-β-D-硫代半乳糖苷(IPTG)诱导6小时和18小时后,PAO1中aiiA基因的表达完全降解了群体感应自诱导物N-酰基高丝氨酸内酯(AHLs):N-氧代十二烷酰-L-高丝氨酸内酯(OdDHL)和N-丁酰-L-高丝氨酸内酯(BHL)。PAO1中AHLs量的减少也与几种毒力因子(如弹性蛋白酶和绿脓菌素)的表达和产生减少相关。AiiA的表达也影响细菌的群体运动性。最重要的是,我们的研究表明,aiiA在铜绿假单胞菌生物膜的形成和分散中起重要作用,这可以通过液体和固体表面上生物膜形成的差异以及扫描电子显微镜下的生物膜结构观察到。