分子伴侣<em>hfq</em>基因缺失对单核细胞增生李斯特菌毒力及生物被膜生成的影响

doi:10.7668/hbnxb.2017.01.015

华北农学报 ›› 2017, Vol. 32 ›› Issue (1): 94-98. doi: 10.7668/hbnxb.2017.01.015

所属专题： 生物技术；

分子伴侣hfq基因缺失对单核细胞增生李斯特菌毒力及生物被膜生成的影响

卢海亭¹, 乔军¹, 孟庆玲¹, 彭叶龙², 陈诚¹, 刘田莉¹, 才学鹏³, 陈创夫¹

1. 石河子大学动物科技学院, 新疆石河子 832003;
2. 阿克苏地区动物疫病控制诊断中心, 新疆阿克苏 843000;
3. 中国农业科学院兰州兽医研究所, 甘肃兰州 730046

收稿日期:2016-11-03 出版日期:2017-02-28
作者简介:卢海亭(1988-),女,河北张家口人,硕士,主要从事病原分子生物学研究。
基金资助:
国家自然科学基金项目（31360596；30960274）；国家国际科技合作专项（No.2014DFR31310）

Effect of Chaperone hfq Gene Deletion on Listeria monocytogenes Virulence and Biofilm Formation

LU Haiting¹, QIAO Jun¹, MENG Qingling¹, PENG Yelong², CHEN Cheng¹, LIU Tianli¹, CAI Xuepeng³, CHEN Chuangfu¹

1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. The Center of Animal Disease Control and Diagnosis in Aksu Area, Aksu 843000, China;
3. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2016-11-03 Published:2017-02-28

摘要/Abstract

摘要： 为了解非编码RNA（non-coding RNA，ncRNA）分子伴侣蛋白Hfq在LM毒力中发挥的调控作用，在构建LM-Δhfq缺失株的基础上，通过动物攻毒试验检测野毒株LM-SB5与缺失株LM-Δhfq对昆明系小鼠的LD₅₀，肝、脾载菌量及对肝、脾、肾的病理损伤；通过溶血试验检测二者的溶血活性；通过结晶紫染色的方法检测二者的生物被膜生成能力。以分析hfq基因缺失对LM毒力及生物被膜生成的影响。结果显示，与LM-SB5相比，LM- Δhfq 对小鼠的LD₅₀升高了6.067倍，毒力显著降低；肝、脾载菌量在第2，4，5天显著下降（P<0.05），在第3天下降极显著（P<0.01）；对肝、脾、肾的病理损伤也有所减弱，LM-Δhfq较LM-SB5溶血能力下降了2倍，生物被膜生成能力在24 h时显著降低（P<0.05）。研究结果证实，Hfq蛋白对LM的毒力及生物被膜生成能力具有重要调控作用。

关键词: 单核细胞增生李斯特菌, 非编码RNA, hfq基因, 毒力

Abstract: To understand the impacts of non-coding RNA (ncRNA) chaprone Hfq protein on LM's virulence,we compared LM- Δhfq with LM-SB5 in the following aspects:LD₅₀to Kunming mouse,amount in mouse liver and spleen as well as effects on pathology of mouse liver,spleen and kidney after inoculation.Hemolysis activity was determined by hemolysis activity assay experiment.Biofilm was prepared and measured to compare the capability of biofilm formation by using crystal violet staining.The results showed that LD₅₀ of LM-Δhfq to Kunming mouse was increased by 6.067 times than that of LM-SB5,indicating strain LM- Δhfq had significantly decreased virulence.LM-Δhfq amount in liver and spleen was significantly lower than that of LM-SB5 in the days of 2, 4, 5 after infection and extremely significant lower in the days of 3 after infection; pathological damages of liver,spleen and kidney of mouse infected LM- Δhfq was declined compared to that of LM-SB5 infection.Hemolysis activity of LM- Δhfq decreased 2 times than that of LM-SB5.The amount of biofilm formed by LM- Δhfq was attenuated.Taking together,the results indicated that ncRNA chaprone Hfq protein is crucial in regulating LM virulence and capability of biofilm formation.

Key words: Listeria monocytogenes, Non-coding RNA, hfq gene, Virulence

中图分类号:

S852

卢海亭, 乔军, 孟庆玲, 彭叶龙, 陈诚, 刘田莉, 才学鹏, 陈创夫. 分子伴侣hfq基因缺失对单核细胞增生李斯特菌毒力及生物被膜生成的影响[J]. 华北农学报, 2017, 32(1): 94-98. doi: 10.7668/hbnxb.2017.01.015.

LU Haiting, QIAO Jun, MENG Qingling, PENG Yelong, CHEN Cheng, LIU Tianli, CAI Xuepeng, CHEN Chuangfu. Effect of Chaperone hfq Gene Deletion on Listeria monocytogenes Virulence and Biofilm Formation[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(1): 94-98. doi: 10.7668/hbnxb.2017.01.015.

http://www.hbnxb.net/CN/Y2017/V32/I1/94

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分子伴侣hfq基因缺失对单核细胞增生李斯特菌毒力及生物被膜生成的影响

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