鸭疫里默氏杆菌ABC外排泵GE296_RS03245基因缺失株的构建及其介导的有机溶剂耐受性  

Construction of ABC efflux pump GE296_RS03245 gene deletion mutant of Riemerella anatipestifer and its contribution to organic solvent resistance

作　　者：朱雨佳 宫晓炜[2] 陈启伟[2] 权衡 秦明星 王燕萍 郑福英[2] 蔺国珍[1] ZHU Yu-jia;GONG Xiao-wei;CHEN Qi-wei;QUAN Heng;QIN Ming-xing;WANG Yan-ping;ZHENG Fu-ying;LIN Guo-zhen(College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China;Stale Key Laboratory of Veterinary Etiological Biology/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,甘肃兰州730030 [2]中国农业科学院,兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2021年第10期1295-1301,共7页Chinese Veterinary Science

基　　金：西北民族大学预防兽医学创新团队项目(110130143);甘肃省自然科学基金项目(21JR1RA203);中国农业科学院中央级公益性科研院所基本科研业务费专项(1610312020001)。

摘　　要：为了明确鸭疫里默氏杆菌RA-LZ01株的GE296RS03245基因(编码ABC外排泵的NBD结构域)在有机溶剂外排中是否发挥作用,本研究利用结合转移和同源重组的方法构建GE296RS03245基因缺失株RAΔ3245,测定亲本株RA-LZ01和基因缺失株RAΔ3245的生长曲线及其对有机溶剂的耐受性,用实时定量RT-PCR测定在有机溶剂刺激下亲本株GE296RS03245基因的转录水平。结果,与亲本株相比,RAΔ3245菌株的生长速率无明显改变,对有机溶剂甲醇、乙醇和异丁醇的敏感性明显升高;在上述有机溶剂刺激下,亲本株GE296RS03245基因的转录水平显著上升。这些结果说明,GE296RS03245基因介导鸭疫里默氏杆菌对有机溶剂甲醇、乙醇和异丁醇的耐受性。In order to determine whether the GE296RS03245 gene(encoding NBD domain of ABC efflux pump) of Riemerella anatipestifer RA-LZ01 strain playing a role in the efflux of organic solvents,GE296RS03245 gene deletion mutant RAΔ3245 was constructed by combining transfer and homologous recombination.The growth curves of the parental strain RA-LZ01 and the deletion mutant RAΔ3245 and the strains’ tolerance to organic solvents were determined.The transcription levels of GE296RS03245 gene of RA-LZ01 strain treated with organic solvent were determined by real-time quantitative RT-PCR.Compared with the parental strain,the growth rate of the deletion mutant RAΔ3245 did not change significantly,but its sensitivity to organic solvents including methanol,ethanol and isobutanol increased significantly.The transcriptional levels of GE296RS03245 gene of parental strain treated with organic solvents increased significantly.These results indicated that GE296RS03245 gene contributed to R.anatipestifer to resist methanol,ethanol and isobutanol.

关 键 词：鸭疫里默氏杆菌 GE296RS03245基因 生长曲线 有机溶剂 

分 类 号：S852.61[农业科学—基础兽医学]