生物学杂志

生物学杂志 ›› 2022, Vol. 39 ›› Issue (1): 27-.doi: 10. 3969/ j. issn. 2095-1736. 2022. 01. 027

• 研究报告 • 上一篇    下一篇

大肠杆菌fliC 基因失活突变株的构建及其特征

  

  1. 1. 贵州医科大学 地方病与少数民族疾病教育部重点实验室,贵阳550004;2. 贵州医科大学 附属医院ICU, 贵阳550004; 3. 青岛大学附属医院 检验科, 青岛266061;4. 贵州医科大学 微生物学教研室, 贵阳550025
  • 出版日期:2022-02-18 发布日期:2022-02-15
  • 通讯作者: 洪伟,博士,教授,主要从事微生物遗传与基因功能研究,E-mail:hongwei@ gmc. edu. cn;齐晓岚,博士,教授,主要从事老年痴呆分子发病机制研究,E-mail:xiaolan76@ 163. com
  • 作者简介:赵行行,硕士研究生,从事微生物遗传与基因功能研究,E-mail:XingxingZhao_56@163.com
  • 基金资助:
    国家自然科学基金项目(U1812403);贵州省自然科学基金项目([2020]1Z067)

Construction and characterization of fliC gene-inactivated mutant of Escherichia coli

  1. 1. Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University, Guiyang550004, China; 2. Department of ICU, The Affiliate Hospital of Guizhou Medical University, Guiyang 550004, China;3. Department of Clinical Laboratory, The Affiliate Hospital of Qingdao University, Qingdao 266061, China;4. Department of Microbiology, Guizhou Medical University, Guiyang 550025, China)
  • Online:2022-02-18 Published:2022-02-15

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摘要：

构建编码大肠杆菌鞭毛蛋白基因(fliC)失活突变株,并探究其生物被膜形成改变后在应激状态下的生物学特征。使用Thermotargetron 靶向基因失活系统,构建fliC 基因失活突变株(ΔfliC420s),测定ΔfliC420s 与野生型菌株的生长速率、运动能力、生物被膜形成能力、自溶速率、pH 敏感性及对抗生素耐受性的变化。ΔfliC420s 相比野生型菌株,生长速率未受影响,在半固体培养基中运动能力缺失,生物被膜形成能力明显降低,菌体自溶速率加快,pH 敏感性增强,以及对D-环丝氨酸、红霉素、诺氟沙星的耐受性降低。大肠杆菌fliC 基因在细菌生物被膜形成和对外界压力的抵抗中具有重要作用。


关键词: 大肠杆菌, fliC 基因, 生物被膜, 基因失活, Thermotargetron

Abstract:

To construct flagellin gene (fliC) inactivation mutant of Escherichia coli (ΔfliC) and explore its biological characteristics under stresses after alteration of biofilm formation. A fliC gene-inactivated mutant strain (ΔfliC420s) was constructed using the Thermotargetron targeted gene inactivation system. The changes of growth rate, motility, biofilm formation ability, autolysis rate, pH sensitivity and antibiotic tolerance of ΔfliC420s and wild-type strain were measured. Compared with wild-type strain, ΔfliC420s mutant had no effect on growth rate, its motility in semi-solid medium was reduced, the biofilm formation ability was significantly reduced, the autolysis rate of bacteria was accelerated, the pH sensitivity was enhanced, Interestingly, the tolerance of ΔfliC420s mutant to D-cycloserine, erythromycin, norfloxacin was reduced. The fliC gene of E. coli plays an important role in the formation of bacterial biofilm and resistance to external stresses.

Key words: Escherichia coli, fliC gene, biofilm, gene inactivation, Thermotargetron

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