过表达哈维弧菌lexA基因对恩诺沙星敏感性的影响
Effect of overexpression of lexA gene of Vibrio harveyi on the sensitivity of enrofloxacin
申晨 1余细红 2王瑞旋 3孙敬锋 1林小植 3朱慧 3郑丽贤 3赖金凤 3梁莉欣3
作者信息
- 1. 天津农学院水产学院,天津 300384
- 2. 揭阳职业技术学院化学工程系,广东揭阳 522000
- 3. 韩山师范学院生命科学与食品工程学院,广东潮州 521041
- 折叠
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.
Abstract
[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.coli β2163(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 enrofloxa-cin.It was confirmed preliminarily that SOS response system was involved in the drug reaction process of V.harveyi to enrofloxacin.
关键词
lexA基因/哈维弧菌/恩诺沙星/SOS反应/耐药传递Key words
lexA gene/V.harveyi/enrofloxacin/SOS response/drug resistance transmission引用本文复制引用
出版年
2024
NETL
NSTL
万方数据