双组分系统resE/resD基因缺失对单增李斯特菌抗渗透压应激能力及致病性的影响

doi:10.16431/j.cnki.1671-7236.2023.09.041

Abstract

Abstract: 【Objective】 This study was aimed to investigate the osmotic stress resistance and pathogenicity of Listeria monocytogenes by constructing a two-component system resE/resD gene deletion strain.【Method】 The two-component system resE/resD gene deletion strain was constructed by homologous recombination.The growth curve assay of Listeria monocytogenes were analyzed to compare the survival performance of the wild strain 10403S and the deletion strain ΔresE/resD.The stress survial test was used to explore the effect of resE/resD gene deletion on the osmotic pressure resistance of Listeria monocytogenes.The effects of resE/resD gene deletion on the cell infiltration ability and pathogenicity of Listeria monocytogenes in mice were evaluated by cell adhesion assay,cell invasion assay and mice virulence assay in colon cancer cells Caco-2 and gastric cancer cells MGC803.【Result】 The growth curve assay results showed that resE/resD gene deletion did not affect the normal growth of Listeria monocytogenes.The growth ability of ΔresE/resD in 5% NaCl was lower than that of wild strain 10403S,and the resistance to osmotic stress in platform stage was significantly decreased (P＜0.05).The results of the stress survival test showed that the survival rate of ΔresE/resD was extremely significantly lower than that of wild strain 10403S after 1 h of stress treatment in 10% NaCl (P＜0.01).In the Caco-2 and MGC803 cell invasion test,the results showed that ΔresE/resD was significantly weaker in cell adhesion (P<0.05),and extremely significantly lower in cell invasion ability (P＜0.01).The results of the mice virulence assay showed that the bacterial load of spleen and liver in mice of ΔresE/resD was extremely significantly or significantly lower than that of wild strain 10403S (P＜0.01 or P＜0.05).【Conclusion】 Two-component system resE/resD gene deletion did not affect the normal growth of Listeria monocytogenes wild strain 10403S,but ΔresE/resD showed reduced resistance to osmotic stress,and weakened the cell adhesion invasiveness and mice pathogenicity of Listeria monocytogenes.

Key words: Listeria monocytogenes; two-component system; resE/resD gene; osmotic stress; pathogenicity

CLC Number:

S852.61⁺7

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GUO Qian, ZHANG Yu, FANG Xiaowei, YUAN Mei, ZHENG Minghao, LIU Fang, FANG Chun. Effects of Two-component System resE/resD Gene Deletion on Osmotic Stress Resistance and Pathogenicity of Listeria monocytogenes[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3853-3860.

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URL: https://www.chvm.net/EN/10.16431/j.cnki.1671-7236.2023.09.041

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Effects of Two-component System resE/resD Gene Deletion on Osmotic Stress Resistance and Pathogenicity of Listeria monocytogenes

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[1]	ZI Jiaxin, LIAN Yingjie, CHEN Minyi, LI Qiuxu, ZHANG Weize, LU Mengke, ZHANG Yangyang, QU Haoxuan, MA Jun, BAI Aiquan. Drug Resistance and Pathogenicity of 4 Strains of Streptococcus suis Type 2 in Guangdong [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3740-3751.
[2]	SHI Chao, ZHANG Wei, LIU Suping, ZHANG Mengqi, HE Xin, ZHAO Mingyan, ZHOU Xia, WANG Zhen, ZHANG Hui. Bioinformatics Analysis and Expression Purification of IspD Protein in Listeria monocytogenes [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3799-3810.
[3]	MA Jing, YANG Danjiao, WEI Longpin, GAN Daibing, LONG Zhixin, HE Yuqi, QING Ting, FAN Yandi, ZHOU Long. Isolation,Identification and Biological Characteristics Analysis of a Moraxella pluranimalium Strain from Tibetan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3275-3285.
[4]	SHENG Xijing, ZHANG Fan, LIU Fangjia, LIU Quan, CAO Zhengzheng, HU Tianyu, LI Donghua, WU Menghui, DAI Menghong. Research Progress on Pathogenicity and Drug Resistance of Klebsiella pneumoniae from Animals [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2966-2973.
[5]	ZENG Dongdong, LIU Caixia, KOU Lijun, QIN He, JIN Ruijie, WANG Jing, REN Jingjing, MA Xun, JIANG Jianjun. Construction and Biological Characteristics of Lm4b_02324 Gene Deletion Strains of Listeria monocytogenes [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1981-1990.
[6]	LIU Caixia, SHI Weidi, KOU Lijun, MA Xun, WANG Jing, GAO Shengjie, LYU Shuangfei, CHEN Xuke, ZENG Dongdong, KONG Cuilian, REN Huijie. Regulation of Transcription of Key Virulence Factors of Listeria monocytogenes by LIPI-4 EⅡC [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1340-1351.
[7]	TIAN Changqing, ZHANG Kunzhong, QI Yumei, SHI Wenjing, DONG Zhijie, ZHANG Haohao, HE Zengwen, ZHI Ji, ZHAO Xuehui, CHONG Qian, XUE Huiwen, GOU Huitian. Isolation,Identification and Biological Characteristics Analysis of a Strain of Listeria monocytogenes from Environment [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1543-1555.
[8]	WANG Lucai, SALANG Wenzhu, ZHANG Huanrong. Isolation,Identification and Drug Resistance and Pathogenicity Analysis of Salmonella Enteritidis from Avian [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1663-1674.
[9]	KANG Yuhui, ZHOU Hongshan, JIN Wenjie, QIN Aijian. Isolation, Identification and Pathogenicity of a Strain of Plesiomonas shigelloides from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3676-3682.
[10]	SHAO Mingzhu, ZHAO Yunqing, LI Songli, HOU Shaohua, ZHANG Yanying. Isolation and Identification of Escherichia coli from Dairy Cows in Hebei Province and Its Pathogenicity and Antibacterial Effect of Chinese Herbal Medicine in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3683-3691.
[11]	WANG Zhifang, XU Yindi, ZHU Wenhao, ZHANG Qingxian, JIAO Wenqiang, LI Haili, WANG Keling. Analysis of Virulence and Drug Resistance of Haemophilus parasuis Serotype 7 Strains [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3245-3255.
[12]	GUO Qian, ZHANG Yu, FANG Xiaowei, AN Ni, YUAN Mei, FANG Chun. Study on Glycosyl-flippase gtcA Gene Deletion Reduces the Pathogenicity of Listeria monocytogenes Serotype 1/2a [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2298-2306.
[13]	WANG Simin, YANG Jiangfeng, ZHAO Xiaokun, ZHANG Lei, GUO Jingjing, LAN Jinping, YU Xiaojie, WANG Jing. Isolation, Identification and Analysis of Pathogenicity and Drug Resistance of Salmonella from Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2385-2396.
[14]	WEI Xiaojun, LI Qiaoqiao, ZENG Dongdong, JIANG Jianjun. Construction of Listeria monocytogenes inlK Gene Deletion Strain and Analysis of Its Biofilm and Disinfectant Resistance [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1870-1878.
[15]	DONG Hongyan, SONG Haigang, ZHU Shanyuan, YU Shengqing, XIE Jun, HONG Shuangpeng, GUO Changming, XU Hai, GAO Yuexiu. Isolation,Identification and Pathogenicity Analysis of Escherichia coli from Ducks [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1977-1984.