食品科学 ›› 2023, Vol. 44 ›› Issue (16): 198-204.doi: 10.7506/spkx1002-6630-20220922-219

• 生物工程 • 上一篇    下一篇

单核细胞增生李斯特氏菌内化素InlJ对噬菌体敏感性及生物被膜形成的影响

刘半红,胡梁斌,陆睿,吴立婷,包红朵,周艳,王冉,张辉   

  1. (1.陕西科技大学食品与生物工程学院,陕西 西安 710021;2.江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地,江苏省农业科学院农产品质量安全与营养研究所,江苏 南京 210014;3.江苏大学食品与生物工程学院,江苏 镇江 212013)
  • 出版日期:2023-08-25 发布日期:2023-09-01
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2018YFE0101900);江苏省农业科技自主创新资金项目(cx(21)1004); 江苏省重点研发计划项目(BE2022361)

Effect of Internalin InlJ of Listeria monocytogenes on Phage Sensitivity and Biofilm Formation

LIU Banhong, HU Liangbin, LU Rui, WU Liting, BAO Hongduo, ZHOU Yan, WANG Ran, ZHANG Hui   

  1. (1. School of Food and Biological Engineering, Shaanxi University of Science and Technology, Xi’an 710021, China; 2. Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Food Safe and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 3. School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China)
  • Online:2023-08-25 Published:2023-09-01

摘要: 为深入探索单核细胞增生李斯特氏菌(Listeria monocytogenes,Lm)inlJ基因在噬菌体敏感性和生物被膜中的作用及功能,本研究通过同源重组构建inlJ基因缺失株Lm NJ05-ΔinlJ,鉴定其生长、黏附及侵袭特性,解析其对噬菌体敏感性和生物被膜形成中的作用机制。结果表明:与野生型Lm NJ05相比,构建的缺失株Lm NJ05-ΔinlJ对RAW264.7细胞的黏附率和侵袭率分别为20.05%和4.42%,黏附和侵袭能力显著减弱;Lm NJ05-ΔinlJ对李斯特菌噬菌体vB-LmoM-NJ05的成斑率提高至2.72 倍;体外裂解分析表明噬菌体vB-LmoM-NJ05对缺失株Lm NJ05-ΔinlJ裂解效果更强;噬菌体效价分别在105 PFU/mL和108 PFU/mL能够完全抑制和清除Lm NJ05-ΔinlJ生物被膜;生物被膜形成相关基因的转录分析表明,噬菌体作用缺失株Lm NJ05-ΔinlJ后,degU、agrA、agrD、luxS、yneA、recA和hpt基因的转录均下调(表达水平趋向于0)。由此可见,inlJ基因的缺失能够增强Lm对噬菌体敏感性,下调Lm对细胞侵袭力和生物被膜形成能力。因此,内化素基因inlJ不仅具有调控Lm自身的作用,还能够调节其对噬菌体的相互作用,为噬菌体的生物防控技术开发和应用奠定基础。

关键词: 单核细胞增生李斯特氏菌;内化素InlJ;噬菌体敏感性;细胞黏附和侵袭;生物被膜

Abstract: In order to explore the role and function of the inlJ gene of Listeria monocytogenes (Lm) in phage sensitivity and biofilm, the inlJ gene-deficient strain Lm NJ05-ΔinlJ was constructed by homologous recombination. The growth, adhesion and invasion characteristics of the defective strain were identified. The results showed that compared with the wide-type strain Lm NJ05, the adhesion and invasion of RAW264.7 cells by Lm NJ05-ΔinlJ were significantly reduced to 20.05% and 4.42%, respectively. The efficiency of plaque formation was enhanced by 2.72 folds and phage vB-LmoM-NJ05 had a stronger lytic activity on Lm NJ05-ΔinlJ. Phage vB-LmoM-NJ05 at titers of 105 and 108 PFU/mL could completely inhibit and remove the biofilm of Lm NJ05-ΔinlJ, respectively. Transcriptional analysis of biofilm formation-related genes showed that the transcriptional levels of the degU, agrA, agrD, luxS, yneA, recA and hpt genes were significantly decreased to nearly zero in the defective strain after interacting with phage vB-LmoM-NJ05. In conclusion, deletion of the inlJ gene can enhance the phage sensitivity of Lm, and down-regulate the ability of cell invasion and biofilm formation. Therefore, the inlJ gene not only regulate Lm but also affect its interaction with phage, which lays a foundation for the development and application of phage biocontrol.

Key words: Listeria monocytogenes; internalin InlJ; phage sensitivity; cell adhesion and invasion; biofilm

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