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

作　　者：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[1] 苟惠天[1] ZHI Ji;CAO Qing;ZHAO Xuehui;ZHANG Haohao;FAN Ziqiu;MA Yonghui;DENG Jing;HE Zengwen;MA Jinrui;ZHANG Kunzhong;CHONG Qian;WANG Caixia;XUE Huiwen;GOU Huitian(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Lintao Vocational and Technical Education Center,Dingri,Gansu 730500,China)

机构地区：[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

出　　处：《中国兽医学报》2024年第9期1923-1929,1956,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31960726,32060822,31560700);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)。

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 H_(2)O_(2)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。This study aims to investigate the function of lmo2363 gene in stress resistance of Listeria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complementation strain of Listeria monocytogenes were constructed using overlapping extended PCR and homologous recombination techniques,and the growth ability,stress survival rate and biofilm formation ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocytogenes 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 H_(2)O_(2)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 biofilm.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 

分 类 号：S852.612[农业科学—基础兽医学]