毒力基因调控蛋白PrfA促进单核细胞增生李斯特菌生物被膜的形成

单核细胞增生李斯特菌(Listeria monocytogenes,LM)是重要的革兰氏阳性食源性致病菌,易在食品以及各种食品加工、运输和保藏设备的接触面形成生物被膜,从而具有更强的抗逆性而难以彻底清除,因此成为食品卫生安全的重要隐患.PrfA是LM毒力基因转录表达的重要调控因子,通过比较研究LM野生株(EGD和EGDe)、PrfA缺失株(EGDAprfA和EGDeAprfA)、无害李斯特菌(Listeria innocua,LI),携带组成性表达PrfA蛋白的重组无害李斯特菌(LI-pERL3-prfA*)以及重组单核细胞增生李斯特菌(EGDeΔprfA-pERL3-prfA*)生物被膜形成能力的差异,探讨LM重要的毒力调控蛋白PrfA对生物被膜形成的影响.实验结果显示:LM野生株具有较强的生物被膜形成能力,而LI形成生物被膜的能力最弱;PrfA的缺失能降低LM生物被膜的形成能力;组成性高量表达PrfA蛋白可以回复EGDeΔprfA的生物被膜形成能力,但对LI没有增强作用.以上实验结果表明:PrfA在LM生物被膜形成中具有重要的促进作用.     

Differential expression of virulence and stress fitness genes during interaction between Listeria monocytogenes and Bifidobacterium longum

Bifidobacterium is well known to have an inhibitory effect on the survival, growth, and proliferation of various foodborne pathogens, but the mechanism of the molecular action of B. longum in blocking the invasion of Listeria monocytogenes is not yet well defined. In the present study, following RNA extraction and cDNA synthesis, differential expression of virulence and stress fitness genes in L. monocytogenes and B. longum was determined by real-time PCR. The results indicate that L. monocytogenes virulence factors, including actA, hly, inlA, and plcA, showed significantly downregulated expression during co-incubation of B. longum and L. monocytogenes in phosphate-buffered saline. The relative mRNA levels of oppA and serpin, two stress fitness genes in B. longum, were significantly higher than for the control group. These results indicate that downregulation of L. monocytogenes virulence factors during co-incubation with B. longum might be responsible for the inhibitory effects.