生物技术通报 ›› 2023, Vol. 39 ›› Issue (4): 288-296.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0917

• 研究报告 • 上一篇    下一篇

恩诺沙星胁迫下嗜水气单胞菌的比较蛋白质组学研究

姚近东1,2,3(), 汤华妹1,2,3, 杨文霄1,2,3, 张丽珊1,2,3, 林向民1,2,3()   

  1. 1.福建农林大学福建省农业生态过程与安全监控重点实验室,福州 350002
    2.福建农林大学作物生态与分子生理重点实验室,福州 350002
    3.福建农林大学生命科学学院,福州 350002
  • 收稿日期:2022-07-24 出版日期:2023-04-26 发布日期:2023-05-16
  • 通讯作者: 林向民,男,博士,教授,研究方向:微生物分子生态学;E-mail:xiangmin@fafu.edu.cn
  • 作者简介:姚近东,男,研究方向 :微生物分子生态学 ;E-mail :945403230@qq.com
    第一联系人:

    汤华妹同为本文第一作者

  • 基金资助:
    :福建省自然科学基金重点项目(2020J02023);省级大学生创新创业计划项目(S202210389069)

Comparative Proteomics Analysis of Aeromonas hydrophila Under Enrofloxacin Stress

YAO Jin-dong1,2,3(), TANG Hua-mei1,2,3, YANG Wen-xiao1,2,3, ZHANG Li-shan1,2,3, LIN Xiang-min1,2,3()   

  1. 1. Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, Fujian Agriculture and Forestry University, Fuzhou 350002
    2. Key Laboratory of Crop Ecology and Molecular Physiology(Fujian Agriculture and Forestry University), Fuzhou 350002
    3. School of Life Sciences(Fujian Agriculture and Forestry University), Fuzhou 350002
  • Received:2022-07-24 Published:2023-04-26 Online:2023-05-16

摘要:

为深入了解喹诺酮类抗生素胁迫下细菌产生的耐药机制,本研究以水生病原菌嗜水气单胞菌为研究对象,利用定量蛋白质组学方法比较嗜水气单胞菌在有无恩诺沙星处理下的蛋白表达差异。质谱结果发现446个差异蛋白,其中233个蛋白表达上调,213个蛋白表达下调。生物信息学分析显示,嗜水气单胞菌可能通过DNA修复和硫代谢相关蛋白的上调表达,促进细菌存活。进一步通过生长曲线测定发现,参与硫代谢的半胱氨酸与恩诺沙星联用能更好地抑制细菌的生长。同时,利用qPCR技术验证硫代谢相关基因在mRNA水平的表达量,发现大部分相关基因在转录水平上的差异表达与蛋白水平一致。以上研究结果表明硫代谢对嗜水气单胞菌在ENR胁迫下起着重要作用。

关键词: 嗜水气单胞菌, 恩诺沙星, LC-MS/MS, 硫代谢, DNA修复

Abstract:

In order to understand the mechanism of bacterial drug resistance under the stress of quinolone antibiotics, this study took Aeromonas hydrophila as the research object, quantitative proteomics was used to compare the protein expression differences in A. hydrophila with and without enrofloxacin treatment. The results showed that total 446 differentially expressed proteins were identified by mass spectrometry, including 233 up-regulated proteins and 213 down-regulated proteins. Bioinformatics analysis showed that A. hydrophila survival may be promoted by up-regulating the expressions of most DNA repair and sulfur metabolism related proteins. Through the growth curve determination, cysteine combined with enrofloxacin can better inhibit the growth of bacteria. Meanwhile, qPCR was used to verify the expression levels of sulfur metabolism-related genes at the mRNA level, and it was uncovered that the differential expression levels of most genes at the transcription level were consistent with the protein level. These results indicate that sulfur metabolism plays an important role in the ENR stress of A. hydrophila.

Key words: Aeromonas hydrophila, enrofloxacin, LC MS/MS, sulfur metabolism, DNA repair