Uhlich Gaylen A, Wonderling Laura D, Luchansky John B
Microbial Food Safety Research Unit, Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, Wyndmoor, Pennsylvania 19038, USA.
Food Microbiol. 2006 May;23(3):300-6. doi: 10.1016/j.fm.2005.03.007. Epub 2005 May 31.
A whole-genome sequence analysis of Listeria monocytogenes strain F2365 revealed 15 potential members of the Crp/Fnr family of transcriptional regulatory proteins. Each gene and the flanking regions were cloned, subjected to in vitro transpositional mutagenesis, and recombined into strain F2365. Mutant strains, produced for 14 of the family members, were compared to strain F2365 for differences in carbon utilization, resistance to oxidative stress, and growth under reduced oxygen conditions that would signal an Fnr- or Crp-like function for these proteins. There were no differences among strain F2365 and the 14 mutant strains in the utilization of the carbon sources readily utilized by L. monocytogenes. Although strain KO2 had a reduced growth rate compared to strain F2365 and the other mutant strains at 30 degrees but not at 37 degrees C, there were no differences in growth rates among strain F2365 and the mutant strains when incubated at either 30 or 37 degrees C under reduced oxygen conditions. However, when compared for differences in response to oxidative stress, mutants KO2 and KO5 showed reduced oxidative stress tolerance compared to the wild-type strain F2365. These results suggest that certain members of the putative Crp/Fnr family in L. monocytogenes may function in response to oxidative stress similar to the Fnr-like protein (Flp) of other gram-positive bacteria.
对单核细胞增生李斯特菌菌株F2365进行的全基因组序列分析揭示了转录调节蛋白Crp/Fnr家族的15个潜在成员。每个基因及其侧翼区域都被克隆出来,进行体外转座诱变,然后重组到菌株F2365中。针对该家族中的14个成员构建了突变菌株,并将其与菌株F2365比较碳利用、抗氧化应激能力以及在低氧条件下的生长情况,以确定这些蛋白质是否具有类似Fnr或Crp的功能。在单核细胞增生李斯特菌易于利用的碳源利用方面,菌株F2365和14个突变菌株之间没有差异。尽管菌株KO2在30℃而非37℃下与菌株F2365和其他突变菌株相比生长速率降低,但在低氧条件下于30℃或37℃培养时,菌株F2365和突变菌株之间的生长速率没有差异。然而,在比较对氧化应激的反应差异时,与野生型菌株F2365相比,突变体KO2和KO5表现出氧化应激耐受性降低。这些结果表明,单核细胞增生李斯特菌中假定的Crp/Fnr家族的某些成员可能在响应氧化应激时发挥作用,类似于其他革兰氏阳性细菌的Fnr样蛋白(Flp)。