Boucher David J, Adler Ben, Boyce John D
Australian Research Council Centre for Structural and Functional Microbial Genomics, Department of Microbiology, Monash University, Victoria 3800, Australia.
J Bacteriol. 2005 Apr;187(7):2278-85. doi: 10.1128/JB.187.7.2278-2285.2005.
Pasteurella multocida is the causative agent of a range of diseases with economic importance in production animals. Many systems have been employed to identify virulence factors of P. multocida, including in vivo expression technology (IVET), signature-tagged mutagenesis, and whole-genome expression profiling. In a previous study in which IVET was used with P. multocida, nrfE was identified as a gene that is preferentially expressed in vivo. In Escherichia coli, nrfE is part of the formate-dependent nitrite reductase system involved in utilizing available nitrite as an electron accepter during growth under anaerobic conditions. In this study, we constructed an isogenic P. multocida strain that was unable to reduce nitrite under either aerobic or anaerobic conditions, thereby demonstrating that P. multocida nrfE is essential for nitrite reduction. However, the nrfE mutant was still virulent in mice. Real-time reverse transcription-PCR analysis indicated that nrfE was regulated independently of nrfABCD by an independent promoter that is likely to be upregulated in vivo.
多杀性巴氏杆菌是引起生产动物一系列具有经济重要性疾病的病原体。许多系统已被用于鉴定多杀性巴氏杆菌的毒力因子,包括体内表达技术(IVET)、标记突变和全基因组表达谱分析。在之前一项使用IVET研究多杀性巴氏杆菌的实验中,nrfE被鉴定为一个在体内优先表达的基因。在大肠杆菌中,nrfE是甲酸依赖型亚硝酸还原酶系统的一部分,该系统在厌氧条件下生长时参与利用可用的亚硝酸盐作为电子受体。在本研究中,我们构建了一株在有氧或无氧条件下均无法还原亚硝酸盐的多杀性巴氏杆菌同基因菌株,从而证明多杀性巴氏杆菌nrfE对于亚硝酸盐还原至关重要。然而,nrfE突变体在小鼠中仍具有毒力。实时逆转录-PCR分析表明,nrfE由一个独立的启动子独立于nrfABCD进行调控,该启动子可能在体内被上调。