金黄色葡萄球菌USA300JE2 sfa、sbn及sfa和sbn基因敲除株构建  

Construction of sfa and/or sbn gene deletion strains of Staphylococcus aureus USA300 JE2

作　　者：赵艳丰[1] 杜冰钰 Simon Heilbronner Zhao Yanfeng;Du Bingyu;Simon Heilbronner(Department of Laboratory Medicine Center,Second Affiliated Hospital of Nanjing Medical University,Nanjing 2100111,China;Department of Microbiology,University of Tübingen,Tübingen 72531,Germany)

机构地区：[1]南京医科大学第二附属医院检验医学中心,南京2100111 [2]德国图宾根大学微生物学系,图宾根72531

出　　处：《中华生物医学工程杂志》2022年第2期155-158,共4页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金(81802071)。

摘　　要：目的构建金黄色葡萄球菌USA300 JE2 sfa和sbn基因敲除株并研究铁载体相关基因对金黄色葡萄球菌生存的影响。方法以pIMAY质粒为载体,构建重组质粒pIMAY-∆sfa、pIMAY-∆sbn,电转入金黄色葡萄球菌USA300 JE2,利用同源重组技术分别构建金黄色葡萄球菌USA300 JE2∆sfa、∆sbn及∆sfa∆sbn基因敲除株,并对基因敲除株进行生长曲线测定。结果通过构建含同源片段的重组质粒,成功敲除sfa基因、sbn基因及sfa和sbn基因,3种敲除株在缺铁条件下的生存能力有不同程度的下降。结论sfa基因和sbn基因敲除对铁限制环境下金黄色葡萄球菌生存有影响。Objective To construct the sfa and/or sbn gene deletion strains of Staphylococcus aureus USA300 JE2,and to investigate the effect of siderophore-related genes on viability of S.aureus.Methods pIMAY plasmid was used as the vector for construction of the recombinant plasmids pIMAY-∆sfa and pIMAY-∆sbn which were subsequently electrotransformed into the S.aureus strain USA300 JE2.Homologous recombination techniques were used to construct the S.aureus USA300JE2 strains with∆sfa,and/or∆sbn knockout.The growth curve of gene knockout strains were measured.Results By constructing recombinant plasmids containing homologous fragments,gene knockout of sfa and/or sbn were successful.The three knockout strains showed varied decline in viability under iron-deficient conditions.Conclusion In iron-limited environments,knockout of sfa and/or sbn gene may affect the viability of Staphylococcus aureus.

关 键 词：金黄色葡萄球菌 USA300 JE2 铁载体 基因敲除 

分 类 号：R378.11[医药卫生—病原生物学]