The Regulatory Roles of ncRNA Rli60 in Adaptability of Listeria monocytogenes to Environmental Stress and Biofilm Formation
The Regulatory Roles of ncRNA Rli60 in Adaptability of Listeria monocytogenes to Environmental Stress and Biofilm Formation
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ncRNA Rli60 在单核细胞增生李斯特氏菌对环境应激和生物膜形成的适应性中的调节作用
DOI：
10.1007/s00284-016-1028-6
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发表时间：
2016-07-01
影响因子：
2.6
通讯作者：
Chen, Chuang-fu
中科院分区：
文献类型：
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作者：
Peng, Ye-long;Meng, Qing-ling;Chen, Chuang-fu
单核细胞增生李斯特氏菌是一种兼性厌氧革兰氏阳性细菌。它能很好地适应外部环境，能够感染人类和动物。为了了解ncRNA Rli60对单增李斯特菌环境胁迫适应性和生物膜形成的影响，采用重叠延伸PCR（SOE-PCR）剪接和同源重组技术构建了单增李斯特菌rli60缺失菌株（LM-Δrli60），并通过测定其在不同温度胁迫下的生长情况，将其与野生型单增李斯特菌EGD-e在环境胁迫适应性方面进行了比较。酸性、碱性、高渗和酒精条件，使用结晶紫染色观察生物膜形成的能力，以及通过实时定量 PCR (qRT-PCR) 观察与生物膜形成相关的基因（gltB 和 gltC）的转录水平。结果表明：（1）在低温（30℃）、高温（42℃）以及碱性和酒精条件下，LM-Δrli60菌株的生长明显减慢；（2）LM-Δrli60形成的生物膜量减弱；（3）LM-Δrli60中gltB和gltC基因在24 h和48 h的转录水平显着降低。减少。总体而言，结果证实 ncRNA Rli60 可能通过影响 gltB 和 gltC 基因的表达，在调节单核细胞增生李斯特氏菌对环境胁迫和生物膜形成的适应性方面发挥重要作用。
Listeria monocytogenes is a facultative anaerobic Gram-positive bacterium. It is well adapted to external environments and able to infect both humans and animals. To understand the impacts of ncRNA Rli60 on the adaptability of L. monocytogenes to environmental stresses and biofilm formation, a rli60 deletion strain of L. monocytogenes (LM-Δrli60) was constructed using splicing by overlap extension PCR (SOE-PCR) and homologous recombination and then compared it with wild-type strain L. monocytogenes EGD-e in the aspects of adaptability to environmental stresses by measuring their growth under stresses of different temperatures, and acidic, alkaline, hypertonic and alcoholic conditions, and capability of biofilm formation by using crystal violet staining, as well as the transcriptional levels of genes (gltB and gltC) related to the biofilm formation by real-time quantitative PCR (qRT-PCR). The results showed that (1) the growth of LM-Δrli60 strain was significantly slower under environmental stresses of low temperature (30 °C), high temperature (42 °C), as well as alkaline and alcoholic conditions, (2) the amount of biofilm formed by LM-Δrli60 was attenuated, and (3) the transcriptional levels of gltB and gltC genes at 24 h and 48 h in LM-Δrli60 revealed a significant reduction. Overall, the results confirmed that ncRNA Rli60 plays important roles in regulating the adaptability of L. monocytogenes to environmental stresses and biofilm formation possibly through impacting the expression of gltB and gltC genes.