Jin Yongxin, Li Tiemei, Xia Huiming, Gu Caifeng, Zhang Xiuming, Bai Yanling, Xu Haijin, Qiao Mingqiang
College of Life Science, Nankai University, China.
Wei Sheng Wu Xue Bao. 2008 Dec;48(12):1623-8.
Pseudomonas aeruginosa is an opportunistic pathogen responsible for a wide range of acute and chronic infections. When growing in the host, it secrets a lot of virulence factors including elastase. This work aimed to explore the genes involved in hydrolyzing ability of Pseudomonas aeruginosa towards elastin.
We performed a transposon mutagenesis analysis of P. aeruginosa PA68 to identify candidate genes involved in elastin hydrolysis. We also monitored the promoter activity of the lasB, a gene encoding the elastase, in the mutants and the wild-type by introducing a Pl(asB-)lacZ transcriptional fusion.
Four mutants with altered levels of elastase production were isolated (elastase activity relative to wild-type was shown in parenthesis): 10 (51%), 17 (131%), 27 (8%) and 84 (13%). Locations of the transposon were mapped to the genome lasA, galU, xcpZ and ptsP, respectively. The results of the lasB promoter's activity were consistent with the elastase activity data (fbetagalactosidase activity relative to wild-type was shown in parenthesis): 10(75%), 17(201%), 27(54%) and 84(7%).
Taken together, the data build up a connection of these four genes with elastase production. This is the first report that gene galU and ptsP may be employed in the regulation of the biosynthesis of elastase.
铜绿假单胞菌是一种机会致病菌,可引发多种急慢性感染。在宿主体内生长时,它会分泌多种毒力因子,包括弹性蛋白酶。本研究旨在探索铜绿假单胞菌中与弹性蛋白水解能力相关的基因。
我们对铜绿假单胞菌PA68进行转座子诱变分析,以鉴定参与弹性蛋白水解的候选基因。通过引入Pl(asB -)lacZ转录融合,我们还监测了编码弹性蛋白酶的lasB基因在突变体和野生型中的启动子活性。
分离出四个弹性蛋白酶产生水平改变的突变体(括号内显示相对于野生型的弹性蛋白酶活性):10(51%)、17(131%)、27(8%)和84(13%)。转座子的位置分别映射到基因组的lasA、galU、xcpZ和ptsP。lasB启动子活性的结果与弹性蛋白酶活性数据一致(括号内显示相对于野生型的β - 半乳糖苷酶活性):10(75%)、17(201%)、27(54%)和84(7%)。
综上所述,这些数据建立了这四个基因与弹性蛋白酶产生之间的联系。这是首次报道galU和ptsP基因可能参与弹性蛋白酶生物合成的调控。