中国兽医学报2024,Vol.44Issue(9) :1923-1929,1956.DOI:10.16303/j.cnki.1005-4545.2024.09.11

单增李斯特菌lmo2363基因缺失株的构建及其生物学特性

Construction and biological characterization of lmo2363 gene deletion strain of Listeria monocytogenes

芝吉 曹青 赵学慧 张浩浩 范子秋 马永辉 邓静 何曾文 马金锐 张坤中 崇倩 王彩霞 薛惠文 苟惠天
中国兽医学报2024,Vol.44Issue(9) :1923-1929,1956.DOI:10.16303/j.cnki.1005-4545.2024.09.11

单增李斯特菌lmo2363基因缺失株的构建及其生物学特性

Construction and biological characterization of lmo2363 gene deletion strain of Listeria monocytogenes

芝吉 1曹青 1赵学慧 1张浩浩 1范子秋 1马永辉 1邓静 1何曾文 1马金锐 1张坤中 1崇倩 1王彩霞 2薛惠文 1苟惠天1
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作者信息

  • 1. 甘肃农业大学动物医学院,甘肃兰州,730070
  • 2. 临洮县职业技术教育中心,甘肃定西 730500
  • 折叠

摘要

探究lmo2363基因在单增李斯特菌分离株LM83-1抗胁迫过程中的功能.本研究以分离株LM83-1为亲本株,利用重叠延伸PCR和同源重组技术,构建单增李斯特菌lmo2363基因缺失株和回补株,比较野生株、缺失株和回补株在不同胁迫环境下的生长能力、应激存活率以及生物被膜形成能力.结果显示,成功构建了单增李斯特菌lmo2363基因缺失株和回补株;生长曲线测定结果显示,在4 ℃、7%NaCl、10%NaCl、3.5%乙醇、4.0%乙醇和pH5胁迫条件下缺失株的生长能力比野生株LM83-1弱(P<0.001);应激存活试验结果表明,在pH3和10 mmol/L H2O2应激处理1 h后,缺失株的存活率显著低于野生株(P<0.010);生物被膜形成能力测定结果显示,与野生株相比,缺失株的生物被膜形成能力下降(P<0.050).结果表明,lmo2363基因介导单核细胞增生性李斯特菌(Listeria monocytogenes,LM)对低温、高渗透压、乙醇和酸胁迫环境的适应以及影响LM生物被膜的形成.本试验结果为进一步探究lmo2363基因在单增李斯特菌抗胁迫过程中的功能奠定了基础.

Abstract

This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7%NaCl,10%NaCl,3.5%ethanol,4.0%ethanol and pH5 stress(P<0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P<0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P<0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

关键词

单增李斯特菌/lmo2363基因/生物被膜/抗胁迫

Key words

Listeria monocytogenes/lmo2363 gene/biofilm/stress resistance

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基金项目

国家自然科学基金资助项目(31960726)

国家自然科学基金资助项目(32060822)

国家自然科学基金资助项目(31560700)

国家重点研发计划资助项目(2019YFC1605705)

甘肃农业大学青年导师扶持基金资助项目(GAU-QDFC-2020-10)

甘肃省重点研发计划资助项目(20YF8FA136)

出版年

2024
中国兽医学报
吉林大学

中国兽医学报

CSTPCDCSCD北大核心
影响因子:0.702
ISSN:1005-4545
参考文献量25
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