Yan An, Wang XiaoLei, Zhang XueHong, Xu YuQuan
Key Laboratory of Microbial Metabolism of Education Ministry, College of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Sci China C Life Sci. 2007 Aug;50(4):518-24. doi: 10.1007/s11427-007-0054-9.
The pltR gene, coding a putative LysR-type regulator, was identified upstream Plt biosynthetic gene cluster in Pseudomonas sp. M18 using bioinformatics technology. The null mutation of pltR resulted in mutant M18TRG (pltR::Gm) by recombination and its Plt (Pyoluteorin) production declined to 30% while PCA (Phenazine-1-carboxylic acid) production remained unchanged as compared with the wild-type M18 grown in King's Medium B. After complementation, Plt production of mutant M18TRG was restored to the level in wild-type M18. Overexpression of pltR in M18 led to 13-fold enhancement of Plt production over the wild-type M18 strain. However, PCA production was unchanged under this condition. These data suggested that PltR was a positive regulator on Plt production. Plt itself, however, could not regulate expression of pltR. Expression of the plt-lacZ transcriptional fusion in mutant M18TRG declined obviously as compared with the wild-type M18, which further proved that PltR could regulate expression of Plt biosynthetic genes at the transcriptional level. In addition, the investigation on the pltR expression in gacA mutant M18G and rsmA mutant M18R disclosed that PltR was involved in the positive regulation of gacA on Plt production while being excluded from the negative control caused by rsmA.
利用生物信息学技术,在假单胞菌属M18中,于吡咯菌素(Plt)生物合成基因簇上游鉴定出编码假定的LysR型调控因子的pltR基因。通过重组,pltR的无效突变产生了突变体M18TRG(pltR::Gm),与在King氏B培养基中生长的野生型M18相比,其吡咯菌素(Plt)产量降至30%,而吩嗪-1-羧酸(PCA)产量保持不变。互补后,突变体M18TRG的Plt产量恢复到野生型M18的水平。在M18中过表达pltR导致Plt产量比野生型M18菌株提高了13倍。然而,在此条件下PCA产量未变。这些数据表明PltR是Plt产量的正调控因子。然而,Plt自身不能调控pltR的表达。与野生型M18相比,突变体M18TRG中plt-lacZ转录融合体的表达明显下降,这进一步证明PltR可在转录水平调控Plt生物合成基因的表达。此外,对gacA突变体M18G和rsmA突变体M18R中pltR表达的研究表明,PltR参与gacA对Plt产量的正调控,而被排除在rsmA引起的负调控之外。