| 摘要: |
| 【目的】构建禽致病性大肠杆菌（APEC）Ⅲ 型分泌系统2（ETT2）转录因子eivF缺失株，并对其进行生物学特性及转录组学分析，探究转录因子eivF基因在APEC致病过程中的作用，为后期深入研究ETT2致病机制奠定基础。【方法】利用Red同源重组技术构建APEC ETT2 转录因子eivF基因缺失的APEC AE81△eivF株及其回复株AE81 C-eivF，通过运动性、生物被膜形成等试验分析eivF对APEC生物学功能的影响；并利用转录组学方法分析eivF在转录调控网络中对细菌鞭毛和生物被膜等相关因子的转录调控作用。【结果】成功构建了APEC eivF基因缺失株AE81△eivF和回复株AE81 C-eivF；生物学结果表明，与野生株AE81相比，缺失株AE81△eivF运动能力降低，生物被膜形成褶皱堆砌且厚度增大，空间结构呈立体、多层次状，并形成类似于三维塔状结构的突起，回复株AE81 C-eivF运动能力和生物被膜形成能力基本恢复；与野生株AE81相比，缺失株AE81△eivF 有576个基因差异表达，且鞭毛和生物被膜等相关基因在转录水平上均发生显著变化。【结论】ETT2的转录因子eivF参与调控APEC的生物表型变化，其可能通过调控鞭毛、生物被膜相关基因而调控APEC致病过程。 |
| 关键词: 禽致病性大肠杆菌 ETT2分泌系统 转录因子eivF |
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| 基金项目:国家自然科学基金项目（31772707）；安徽农业大学2019年度研究生创新基金（2019ysj-24） |
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| Biological characteristics and transcriptomics of avian pathogenic Escherichia coli ETT2 transcription factor eivF deletion strain |
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FU Dandan,XIAO Yating,XUE Mei,et al
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| Abstract: |
| 【Objective】This study constructed avian pathogenic Escherichia coli (APEC) type III secretion system 2 (ETT2) transcription factor eivF deletion strain,analyzed its biological characteristics and transcriptomics,and explored the role of transcription factor eivF gene in APEC2.【Method】The APEC ETT2 transcription factor eivF gene deleted APEC AE81△eivF strain and the complemented strain AE81 C-eivF were constructed by Red homologous recombination technique.The effects of APEC on the biological function of APEC were analyzed by detecting motility and biofilm formation.The transcriptomics analysis was used to analyze the transcriptional regulation of EivF on bacterial flagella and biofilms in transcriptional regulatory networks.【Result】The APEC eivF gene deletion strain AE81△eivF and the complemented strain AE81 C-eivF were successfully constructed.Compared with the wild strain AE81,the mobility of AE81△eivF was reduced,and the biofilm formed folds and its thickness increased.The spatial structure was three-dimensional and multi-layered with a protrusion formed similar to the three-dimensional tower structure.The recovery ability of the AE81 C-eivF and the biofilm formation ability of the complemented strain AE81 were basically restored.Compared to the wild strain AE81,the deletion strain AE81ΔeivF had 576 genes that were differentially expressed,and related genes such as flagella and biofilm were significantly changed at the transcriptional level.【Conclusion】The transcription factor EivF of ETT2 involved in the regulation of the biophenotypic changes of APEC,and it may regulate the pathogenesis of APEC by regulating flagella and biofilm related genes. |
| Key words: avian pathogenic Escherichia coli type Ⅲ secretion system 2 transcription factor eivF |
