摘要
目的探究单核细胞增生李斯特菌生物被膜相关非编码小RNA 00085(sRNA 00085)对该菌生存及环境适应能力的调控作用。方法采用同源重组技术构建sRNA 00085靶基因的缺失株ΔsRNA 00085及回补株CΔsRNA 00085,并对比标准株、缺失株及回补株生长能力、生物被膜形成能力及抗生素敏感性方面的差异。结果与标准株相比,ΔsRNA 00085株的生物被膜形成能力及对青霉素、哌拉西林、强力霉素、四环素、万古霉素、复方新诺明的敏感性显著降低,其中仅对四环素的最低抑菌浓度(MIC)表现出显著降低,而CΔsRNA 00085株在这些特性上与标准株无显著差异。进一步检测发现,敲除sRNA 00085后,单核细胞增生李斯特菌四环素耐药相关基因tetS转录水平显著升高,但对tetA的转录无影响。结论sRNA 00085通过调控单核细胞增生李斯特菌的生物被膜形成能力及多种抗生素耐药性,从而影响其环境适应性。
The purpose of this study was to investigate the regulatory effects of biofilm associated non-coding small RNA(sRNA)00085 on the survival and environmental fitness of Listeria monocytogenes.Homologous recombination technology was used to construct a deletion mutant strain(ΔsRNA 00085)and a complementary strain(CΔsRNA 00085)of the sRNA00085 target gene.The differences in biological characteristics were compared among the standard strain,ΔsRNA 00085,and CΔsRNA 00085.The deletion of sRNA 00085 led to a significant decrease in biofilm formation capacity and sensitivity to several antibiotics,including penicillin,piperacillin,doxycycline,tetracycline,vancomycin,and cotrimoxazole.However,only the minimum inhibitory concentration(MIC)of tetracycline exhibited a significant decrease inΔsRNA 00085.Meanwhile,the decreased biofilm formation and antibiotic resistance of the sRNA 00085 mutant were restored in the CΔsRNA 00085 strain.Furthermore,we investigated the transcription levels of tetracycline resistance-related genes in L.monocytogenes.Down-regulated transcription of the tetS gene but no significant difference in transcription of the tetA gene were observed inΔsRNA 00085 compared with the standard strain and CΔsRNA 00085.Moreover,the elimination of sRNA 00085 did not affect bacterial growth ability or sensitivity to disinfectants.These findings highlight that sRNA 00085 plays an important role in the environmental adaptability of L.monocytogenes by affecting bacterial biofilm formation and resistance.
作者
邓静
芝吉
范子秋
赵学慧
宋雅丽
苟惠天
魏衍全
曹青
DENG Jing;ZHI Ji;FAN Zi-qiu;ZHAO Xue-hui;SONG Ya-li;GOU Hui-tian;WEI Yan-quan;CAO Qing(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2024年第7期620-627,共8页
Chinese Journal of Zoonoses
基金
甘肃农业大学公招博士科研启动金基金项目(No.GAU-KYQD-2022-36)
国家自然科学基金项目(No.32060822,No.31560700)
甘肃省技术创新引导计划项目(No.23CXNA0016)
甘肃省自然科学基金青年项目(No.21JR7RA482)
张家川揭榜挂帅项目(No.ZC-STK-2023A-030)。
