Shao Huifeng, Zhang Bin, Wang Lv, Jiao Haitao, Tang Li, Chen Xuelan
Wei Sheng Wu Xue Bao. 2014 Jun 4;54(6):635-40.
The FarR protein was involved in the regulation of arginine biosynthetic pathway in corynebacterium, but the regulation mechanism of FarR protein and its relationship with the negative regulator ArgR have never been reported. In this work, we constructed two deletion mutants: C. crenatum delta farR and C. crenatum delta argR delta farR, and investigated the FarR function and its relationship with ArgR through the determination of transcriptional levels of arginine biosynthetic genes in four strains, including C. crenatum delta argR constructed in previous work.
We used marker-less knockout technology to construct C. crenatum delta farR and C. crenatum delta argR delta farR, and compared the transcriptional levels of the arginine biosynthetic genes in three mutant strains with those of the wild type strain using real-time fluorescence quantitative PCR.
The results of RT-qPCR indicate that, in the absence of ArgR, FarR acted as a positive regulator. When farR gene was knockout alone, the transcriptional levels of arginine biosynthetic genes appeared up-regulated, down-regulated or no influence.
FarR and ArgR are involved together in the regulation of arginine biosynthetic pathway of C. crenatum.
FarR蛋白参与了棒状杆菌中精氨酸生物合成途径的调控，但FarR蛋白的调控机制及其与负调控因子ArgR的关系尚未见报道。在本研究中，我们构建了两个缺失突变体：钝齿棒杆菌ΔfarR和钝齿棒杆菌ΔargRΔfarR，并通过测定包括前期构建的钝齿棒杆菌ΔargR在内的四株菌中精氨酸生物合成基因的转录水平，研究了FarR的功能及其与ArgR的关系。
我们采用无标记敲除技术构建钝齿棒杆菌ΔfarR和钝齿棒杆菌ΔargRΔfarR，并使用实时荧光定量PCR比较了三株突变体菌株与野生型菌株中精氨酸生物合成基因的转录水平。
RT-qPCR结果表明，在没有ArgR的情况下，FarR作为正调控因子发挥作用。单独敲除farR基因时，精氨酸生物合成基因的转录水平出现上调、下调或无影响。
FarR和ArgR共同参与钝齿棒杆菌精氨酸生物合成途径的调控。
