Department of Food Science, University of Massachusetts, Amherst, MA 01003, USA.
Int J Food Microbiol. 2013 Feb 15;161(3):158-63. doi: 10.1016/j.ijfoodmicro.2012.12.003. Epub 2012 Dec 20.
The distribution and survival of Listeria monocytogenes (L. monocytogenes) in food processing environment is linked to its ability to form biofilms, however the genetic mechanisms remain unclear. In our previous study, a Himar1 mariner-based transposon mutagenesis was performed and 42 mutants were confirmed to have reduced biofilm formation. Among the 42 biofilm deficient mutants, two isolates (s25-10C and s55-1D) harbored single insertion in lmo1386, a gene encoding a putative DNA translocase. The lmo1386 mutants had impaired biofilm formation in both static and flow conditions. The mutant strain s55-1D was complemented by cloning the entire lmo1386 gene into pPL2-gtcAP, a derivative of the integration vector pPL2 with the L. monocytogenes gtcA promoter. The genetically complemented mutant restored its biofilm phenotype, demonstrating the role of lmo1386 in the biofilm formation of L. monocytogenes. The lmo1386 mutant had reduced initial adhesion ability, which could at least partially contribute to the impaired biofilm phenotype. Additionally, the lmo1386 mutant formed elongated cell chains when grown in a nutrient TSBYE media, while no obvious cell morphology changes were observed when grown in the minimal MWB media. Overall, our findings suggest that the disruption of lmo1386, a putative DNA translocase gene affects the biofilm formation of L. monocytogenes on abiotic surfaces, which may further advance the understanding of the complicated process of biofilm formation.
李斯特菌(Listeria monocytogenes)在食品加工环境中的分布和存活与其形成生物膜的能力有关,但遗传机制尚不清楚。在我们之前的研究中,进行了基于 Himar1 转座子的转座子诱变,证实了 42 个突变体生物膜形成能力降低。在 42 个生物膜缺陷突变体中,两个分离株(s25-10C 和 s55-1D)在 lmo1386 中存在单个插入,该基因编码一种假定的 DNA 转位酶。lmo1386 突变体在静态和流动条件下的生物膜形成能力都受到了损害。通过将整个 lmo1386 基因克隆到 pPL2-gtcAP 中,对突变菌株 s55-1D 进行了遗传互补,pPL2-gtcAP 是整合载体 pPL2 的衍生物,带有李斯特菌 gtcA 启动子。遗传互补突变体恢复了其生物膜表型,证明了 lmo1386 在李斯特菌生物膜形成中的作用。lmo1386 突变体初始粘附能力降低,这至少部分导致了其生物膜表型受损。此外,lmo1386 突变体在营养丰富的 TSBYE 培养基中生长时形成伸长的细胞链,而在基本的 MWB 培养基中生长时则没有观察到明显的细胞形态变化。总的来说,我们的研究结果表明,破坏假定的 DNA 转位酶基因 lmo1386 会影响李斯特菌在非生物表面上形成生物膜,这可能进一步加深我们对生物膜形成这一复杂过程的理解。