通过使用CRISPRi系统向RBSB基因来抑制大肠杆菌中的等离子体结合
Yawen Xiao1, Yan Zhang1, Fengjun Xie1
1Institute of Food Safety and Nutrition, Jinan University, Guangzhou 510632, China.
International journal of molecular sciences
|July 14, 2023
概括
用CRISPR干扰准rbsB基因有效地抑制了细菌的结合和抗生素耐药基因的传播. 这种方法减少了Escherichia coli中的等离子体转移,定数感应和生物膜形成.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
背景情况:
- 细菌结合是传播抗生素耐药基因 (ARG) 在病原体之间的主要机制.
- 干扰结合可以限制ARGs的传播.
研究的目的:
- 研究利用CRISPR干扰 (CRISPRi) 系统向rbsB基因的潜力,以抑制大肠杆菌中的等离体介导水平基因转移.
- 评估rbsB基因抑制对结合效率,定数感应和生物膜形成的影响.
主要方法:
- 设计了三种单导向RNA (sgRNA) 来准rbsB基因.
- 在大肠杆菌中构建了一个CRISPRi系统来抑制rbsB表达.
- 量化了rbsB的转录水平,与结合相关的基因和定数感应信号.
- 评估结合效率和生物膜形成率.
主要成果:
- 克里斯普里系统有效地抑制了66.4%的rbsB基因转录.
- 抑制rbsB显著降低了88.7%的RP4等离子体结合,并抑制了关键的结合基因 (trbBp,trfAp,traF,traJ).
- 减少rbsB表达减少了12.8%的自诱导体2 (AI-2) 枯竭和68.2%的生物膜形成.
结论:
- rbsB基因作为一种可行的普遍标,可以抑制细菌的结合.
- 开发的CRISPRi系统显示了在高风险环境中控制ARG传播的潜力.
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