鼠伤寒沙门菌ompW基因缺失株与回补株的构建及其生物学特性  

Construction and Biological Characterization of ompW Gene Deletion and Complemented Strain of Salmonella Typhimurium

作　　者：葛雷 仇汝龙 范志宇[1,2,3] 胡波 魏后军[1,2,3] 陈萌萌 宋艳华[1,2,3] 李一鸣 徐为中[1,2,3] 王芳 GE Lei;QIU Rulong;FAN Zhiyu;HU Bo;WEI Houjun;CHEN Mengmeng;SONG Yanhua;LI Yiming;XU Weizhong;WANG Fang(Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Key Laboratory for Veterinary Bio-Product Engineering,Ministry of Agriculture,Nanjing 210014,China;Guotai(Taizhou)Center of Technology Innovation for Veterinary Biologicals,Taizhou 225300,China)

机构地区：[1]江苏省农业科学院兽医研究所,南京210014 [2]农业农村部兽用生物制品工程重点实验室,南京210014 [3]兽用生物制品(泰州)国泰技术创新中心,泰州225300

出　　处：《畜牧兽医学报》2025年第4期1865-1875,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：现代农业产业技术体系建设专项资金(CARS-43-C-1)。

摘　　要：鼠伤寒沙门菌(Salmonella Typhimurium)是最为常见的沙门菌血清型,严重危害畜牧业发展及公共卫生安全。外膜蛋白OmpW是由ompW基因编码的营养物质特异性运输通道,发挥特殊的铁离子外排功能,与细菌耐药性、致病性等密切相关。但其在沙门菌中的功能尚未见研究。为探究外膜蛋白OmpW对鼠伤寒沙门菌的影响,本试验以鼠伤寒沙门菌ATCC 14028为野生株(WT14028),利用λ-Red同源重组技术构建ompW基因缺失株(14028ΔompW),同时构建回补株(14028ΔompW::ompW),通过检测生长特性、生物被膜形成能力、环境耐受性、耐药性等方面探究鼠伤寒沙门菌ompW基因的生物学特性,并通过小鼠感染试验探究该基因对沙门菌致病性的影响。结果表明,缺失株与野生株菌落形态无显著差异;缺失株在对数生长期显著慢于野生株,但到达稳定期无显著差异;缺失株生物被膜形成能力下降31.73%±6.43%;在酸性(pH=3)、碱性(pH=10)及高盐(1 mol·L^(-1)NaCl)环境下,缺失株存活率显著低于野生株;但在高铁(>0.625 g·L^(-1)FeCl3)环境下,缺失株存活率更高;缺失株对氨基糖苷类抗生素庆大霉素、卡那霉素的敏感性显著增强,对其他如氨苄西林、四环素等抗生素敏感性无显著变化;小鼠感染试验表明,与野生株相比,缺失株感染诱发的腹泻率更低、脾肿大及肠道损伤更轻微,且细菌移位能力也显著下降。综上,本研究成功构建了鼠伤寒沙门菌ompW基因缺失株及回补株,试验表明该基因与菌株生长、环境耐受性、耐药性、致病性等方面密切相关。本研究为进一步开展沙门菌的致病与耐药机制的研究奠定了基础。Salmonella Typhimurium is the most common serotype of Salmonella,which seriously endangers the development of animal husbandry and public health safety.Outer membrance protein OmpW,encoded by ompW,is a special nutrient transport channel that has iron ion efflux function and is closely related to drug resistance,pathogenicity and other factors.However,the function of OmpW in Salmonella has not been studied yet.To investigate the effect of OmpW in Salmonella Typhimurium,this experiment used Salmonella Typhimurium ATCC 14028(WT14028)as a wild strain,constructed ompW gene deletion strain(14028ΔompW)usingλ-Red homologous recombination technology and constructed complemented strain(14028ΔompW::ompW).Growth characteristics,biofilm formation ability,environmental tolerance and antimi-crobial resistance were detected to investigate the biological characteristics of ompW.And the impact on the pathogenicity was explored through mouse infection experiments.The results showed that there was no significant difference in colony morphology between the deletion strain and the wild strain.The deletion strain showed significantly slower in logarithmic growth period than the wild strain,but there is no significant difference in stable period.The biofilm formation ability of the deletion strain decreased by 31.73%±6.43%.In acidic(pH=3),alkaline(pH=10)and high salt(1 mol·L^(-1) NaCl)environments,the survival rate of the deletion strain was significantly lower than that of the wild strain.But in high iron(>0.625 g·L^(-1) FeCl3)environ-ment,the survival rate of the deletion strain was higher.The sensitivity of the deletion strain to aminoglycoside antibiotics such as gentamicin and kanamycin was significantly enhanced,but there was no significant change in sensitivity to other antibiotics such as ampicillin and tetracy-cline.The results of the mouse infection experiments showed that the deletion strain induced lower diarrhea rate,milder splenomegaly and intestinal damage compared with the wild strain.And the bacterial translocatio

关 键 词：鼠伤寒沙门菌 ompW基因 缺失株 生物学特性 致病性 

分 类 号：S852.61[农业科学—基础兽医学]