鼠伤寒沙门菌孔蛋白ompW、ompS、ompD基因突变株的构建及生物学特性分析  

Construction and biological characterization of pore protein ompW,ompS and ompD gene mutant strains of Salmonella typhimurium

作　　者：武绍碧 令狐远凤 潘永 杨婉 陈世雄 叶景芬 杨琦[1,2,4] WU Shaobi;LINGHU Yuanfeng;PAN Yong;YANG Wan;CHEN Shixiong;YE Jingfen;YANG Qi(College of Animal Science,Guizhou University,Guiyang 550025,China;Institute of Animal Diseases,Guizhou University,Guiyang 550025,China;Institute of Animal Husbandry and Veterinary Medicine,Guizhou Academy of Agricultural Sciences,Guiyang 550025,China;Guizhou Animal Disease Research Office,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州大学动物疫病研究所,贵州贵阳550025 [3]贵州省农业科学院畜牧兽医研究所,贵州贵阳550025 [4]贵州省动物疫病研究室,贵州贵阳550025

出　　处：《中国兽医学报》2024年第6期1165-1174,共10页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32260884)。

摘　　要：为探究孔蛋白基因ompW、ompS、ompD对鼠伤寒沙门菌的生物学特性及毒力的影响,本研究利用λ-Red同源重组系统构建相应突变株,通过检测生长曲线、泳动性、生化特性、体外遗传稳定性、生物被膜形成能力、耐药性、以及半数致死量(LD_(50))来比较鼠伤寒沙门菌标准菌株与各突变株之间的区别。结果显示,相比标准株,ompD和ompW基因突变株对细菌生长速率和运动能力影响较小,而ompS基因突变株的生长速率和运动能力则明显降低;3个突变株均不影响鼠伤寒沙门菌的生化特性,也不影响遗传稳定性,但能不同程度影响其对多种常用抗生素的敏感性,且均导致成膜能力极显著下降(P<0.01);LD_(50)毒力测定中,3个突变株毒力均导致鼠伤寒沙门菌的毒力下降,其中ompS基因突变株毒力下降最明显,LD_(50)为标准菌株的25倍。综上所述,孔蛋白ompW、ompS、ompD基因突变能影响鼠伤寒沙门菌的部分生物学特性。本研究结果为进一步研究孔蛋白ompW、ompS、ompD基因的生物学功能及沙门菌致病性奠定了试验基础。In order to investigate the effects of porin genes ompW,ompS and ompD on the biological properties and virulence of Salmonella typhimurium,the corresponding mutant strains were constructed using theλ-Red homologous recombination system,and the growth curves,motility,biochemical properties,in vitro genetic stability,biofilm-forming ability,drug resistance,and lethal dose at half capacity(LD_(50))between the standard strain and each mutant strain were detected by comparative assays for Salmonella typhimurium.The results showed that,compared with the standard strain,the ompD and ompW mutation had less effect on the growth rate and motility of the bacteria,while the ompS mutation significantly reduced the growth rate and motility;none of the three genetic mutation affected the biochemical characteristics of Salmonella typhimurium,nor the genetic stability,but affected its susceptibility to a variety of commonly used antibiotics to varying degrees and caused a highly significant decrease(P<0.01)in the ability to form a biofilm,and the results showed that the three mutant strains had a significant reduction in the ability to form a biofilm.The result of LD_(50)virulence assay showed that all three genetic mutation led to a decrease in the virulence of Salmonella typhimurium,among which the ompS mutant strain showed the most obvious decrease in virulenc,LD_(50)was 25 times that of the standard strain.In conclusion,mutations of the pore protein ompW,ompS,and ompD genes can affect some biological properties of Salmonella typhimurium.The results of this study laid an experimental foundation for further research on the biological functions of the pore protein ompW,ompS and ompD genes and Salmonella pathogenicity.

关 键 词：鼠伤寒沙门菌 孔蛋白 λ-Red同源重组系统 生物学特性 

分 类 号：S852.61[农业科学—基础兽医学]