RaeR对鸭疫里默氏杆菌外排泵RaeC-RaeA-RaeB的调控作用研究  

Study on the Role of RaeR in the Regulation of Efflux Pump RaeC-RaeA-RaeB of Rimerella anatipestifer

作　　者：李倩倩 林馨 韦依琳 崔昊宇 邹荣华 吴晓妮 葛家振 黄国梁 张丽娟 郑福英[2] 蔺国珍[1] LI Qianqian;LIN Xin;WEI Yilin;CUI Haoyu;ZOU Ronghua;WU Xiaoni;GE Jiazhen;HUANG Guoliang;ZHANG Lijuan;ZHENG Fuying;LIN Guozhen(College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China;State Key Laboratory for Animal Disease Control and Prevention,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]西北民族大学生命科学与工程学院,兰州730030 [2]中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室,兰州730046

出　　处：《畜牧兽医学报》2023年第9期3793-3802,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(32260882);中央高校基本科研业务费专项资金(31920220137)。

摘　　要：旨在阐明鸭疫里默氏杆菌RA-LZ01株GE 296_RS05175基因编码的RaeR对外排泵RaeC-RaeA-RaeB的调控作用。构建RA-LZ01株的GE296_RS05160基因(编码内膜蛋白RaeB)缺失株ΔRaeB和回复株cΔRaeB,以及GE296_RS05175基因缺失株ΔRaeR和回复株cΔRaeR。测定菌株的生长曲线、菌株对抗菌素的敏感性以及GE296_RS05160和GE296_RS05175基因在特异性底物刺激下的转录水平。结果显示,与亲本株相比,缺失株的生长能力无明显变化;与亲本株和回复株相比,缺失株ΔRaeB对卡那霉素、链霉素和十二烷基硫酸钠(sodium dodecyl sulfate,SDS)的敏感性上升,而缺失株ΔRaeR对卡那霉素、链霉素和SDS的敏感性降低;加入SDS刺激后,RA-LZ01和ΔRaeR株的GE296_RS05160基因的相对转录水平分别上升了2.26倍和4.64倍,ΔRaeR株中该基因的上调倍数明显超过了RA-LZ01株;在SDS刺激下,RA-LZ01和ΔRaeB株的GE296_RS05175基因的相对转录水平均明显下调。上述结果表明,外排泵RaeC-RaeA-RaeB与菌株的生长无明显相关性,可介导菌株对卡那霉素、链霉素和SDS产生耐受;底物SDS可刺激GE296_RS05175基因的转录水平降低,该基因编码的RaeR对RaeB的表达发挥负向调控作用。The aim of this study was to clarify the role of RaeR encoded by GE296_RS05175 gene in the regulation of efflux pump RaeC-RaeA-RaeB of Rimerella anatipestifer RA-LZ01 strain.The GE296_RS05160 gene(encoding the inner membrane protein RaeB)deletion mutantΔRaeB and complemented strain cΔRaeB of RA-LZ01 strain were constructed.Similarly,GE 296_RS05175 gene deletion mutantΔRaeR and its complemented strain cΔRaeR were constructed.The growth curves,antimicrobial susceptibility and transcription levels of GE296_RS05160 and GE296_RS05175 genes under specific substrate stimulation were measured.Compared with the parental strain,the growth abilities of the deletion mutants had no obvious change.By comparison with the parental strain and complemented strains,the sensitivity ofΔRaeB to kanamycin,streptomycin and(sodium dodecyl sulfate,SDS)were increased,whileΔRaeR showed the opposite results.After SDS stimulation,the relative transcription levels of GE296_RS05160 gene in RA-LZ01 andΔRaeR strains increased 2.26 and 4.64 times,respectively.The upregulation of GE296_RS05160 gene inΔRaeR strain was significantly higher than that in RA-LZ01 strain.Under SDS stimulation,the relative transcription levels of GE296_RS05175 gene in RA-LZ01 andΔRaeB strains decreased significantly.These results indicated that the efflux pump RaeC-RaeA-RaeB had no significant correlation with the growth of RA-LZ01 strain,but could mediate the tolerance of the strain to kanamycin,streptomycin and SDS.The transcription level of GE296_RS05175 gene reduced under SDS pressure,and RaeR,which encoded by GE296_RS05175 gene,regulated the expression of RaeB negatively.

关 键 词：鸭疫里默氏杆菌 RAEB RaeR 调控 

分 类 号：S852.6[农业科学—基础兽医学]