Paterson G K, Cone D B, Peters S E, Maskell D J
Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK.
Microb Pathog. 2009 May;46(5):261-5. doi: 10.1016/j.micpath.2009.01.008. Epub 2009 Feb 5.
To assess the role of the glycolytic enzyme phosphofructokinase (Pfk) in the in vivo fitness of the pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) we have generated single and double gene deletion mutants of the two known isoforms of this enzyme, pfkA and pfkB. In a mouse model of typhoid fever, bacterial counts in the spleen and liver were similar between wild type and single pfkA and pfkB mutant-infected mice. However, a double pfkAB mutant was significantly attenuated for growth in vivo. This defect was complemented by provision of either pfkA or pfkB on pBR322. Together these data show that Pfk activity is required for the full in vivo fitness of S. Typhimurium with functional redundancy between pfkA and pfkB. The level of attenuation of the pfkAB double mutant was not sufficient for its consideration as a live attenuated vaccine strain.
为了评估糖酵解酶磷酸果糖激酶(Pfk)在肠炎沙门氏菌鼠伤寒血清型(鼠伤寒沙门氏菌)体内适应性中的作用,我们构建了该酶两种已知同工型pfkA和pfkB的单基因和双基因缺失突变体。在伤寒热小鼠模型中,野生型以及感染单pfkA和pfkB突变体的小鼠脾脏和肝脏中的细菌计数相似。然而,pfkAB双突变体在体内生长时显著减毒。通过在pBR322上提供pfkA或pfkB可弥补这一缺陷。这些数据共同表明,Pfk活性是鼠伤寒沙门氏菌完全体内适应性所必需的,且pfkA和pfkB之间存在功能冗余。pfkAB双突变体的减毒水平不足以使其被考虑作为减毒活疫苗株。