摘要
[目的]构建过表达lexA基因的哈维弧菌株,研究SOS应答系统与哈维弧菌对恩诺沙星敏感性的关联作用,并对相关耐药机制进行初步的分析。[方法]构建了lexA重组表达质粒pBAD33cm-rp4-lexA,转入大肠杆菌β2163(DH5α)构建重组质粒菌株β2163(pBAD33cm-rp4-lexA),并通过纸片扩散法检测哈维弧菌对恩诺沙星的敏感性变化。[结果]lexA基因过表达菌株构建成功,药敏试验结果显示:哈维弧菌耐药株抑菌圈直径由9 mm变为11 mm,耐药性由耐药变为中度敏感。[结论]成功构建lexA基因过表达菌株,提高了哈维弧菌对恩诺沙星的敏感性,明确了lexA基因与哈维弧菌对恩诺沙星的敏感性直接相关,初步证实了SOS应答系统参与哈维弧菌对恩诺沙星的药物反应过程。
[Objective]Overexpressing lexA gene in Vibrio harveyi strain was constructed to study correlation between the SOS response system and the sensitivity of V.harveyi to enrofloxacin,and related drug resistance mechanism was analyzed prelimina-rily.[Method]Recombinant expression plasmid of pBAD33cm-rp4-lexA was constructed by inserting the lexA gene into the prokaryotic expression vector pBAD33cm-rp4 and transformed into the bacterial competent E.coliβ2163(DH5α)cells namesβ2163(pBAD33cm-rp4-lexA)strain,and sensitivity of V.harveyi to enrofloxacin was detected by Kirby-Bauer method.[Result]overexpression lexA gene strain was constructed successfully.Results of drug sensitivity test showed that the diameter of the inhibition zone of the drug-resistant strain of V.harveyi from 9 mm to 11 mm,and the drug resistance from drug resist-ance to moderate sensitivity.[Conslusion]Overexpression lexA gene strain was successfully constructed,and improved the sensi-tivity of V.harveyi to enrofloxacin.It was confirmed that lexA gene was related directly to the sensitivity of V.harveyi to enroflox-acin.It was confirmed preliminarily that SOS response system was involved in the drug reaction process of V.harveyi to enroflox-acin.
作者
申晨
余细红
王瑞旋
孙敬锋
林小植
朱慧
郑丽贤
赖金凤
梁莉欣
SHEN Chen;YU Xihong;WANG Ruixuan;SUN Jingfeng;LIN Xiaozhi;ZHU Hui;ZHENG Lixian;LAI Jinfeng;LIANG Lixin(College of Fisheries,Tianjin Agricultural University,Tianjin 300384;College of Life Sciences and College of Food Engineering,Hanshan Normal University,Chaozhou 521041;Department of Chemical Engineering,Jieyang Vocational and Technical College,Jieyang 522000,China)
出处
《生物技术》
CAS
2024年第6期671-676,共6页
Biotechnology
基金
广东省重点建设学科科研能力提升项目(2021ZDJS04)
广东省高校重点领域专项(2022ZDZX4029)
国家自然科学基金项目(31902416)
潮州市科技专项(202201NY01)
韩山师范学院校级博士启动项目(QD202119)
“广东省粤东药食资源功能物质与治未病研究重点实验室”开放基金课题项目(GPKLFSHP202302)。
