Construction and Characterization of an lpxM-Deficient Acinetobacter baumannii Strain Using a pyrF/5-FOA Counterselection System

利用pyrF/5-FOA反向选择系统构建和鉴定lpxM缺陷型鲍曼不动杆菌菌株

阅读:1

Abstract

AIM: Multidrug-resistant Acinetobacter baumannii (MDR-AB) is on the rise, making it challenging to achieve the desired therapeutic effects with existing conventional antibiotics. The search for new antibacterial targets has emerged as a significant research focus. PURPOSE: The lysophospholipid acyltransferases (LPLATs) proteins encoded by the lpxM gene play a pivotal role in the biosynthesis of lipopolysaccharides (LPS). LPS is a critical component of the outer membrane of the cell wall and is essential for the survival and drug resistance of Gram-negative bacteria. This study aims to investigate the effects of the lpxM gene on the growth and drug susceptibility of MDR-AB. METHODS: The standard strain of Acinetobacter baumannii (A. baumannii, AB) AYE was selected as the target. The lpxM gene was knocked out using the pyrF/5-FOA-based counterselectable method. Subsequently, the growth status and the minimum inhibitory concentration (MIC) of the knockout strain against conventional antibiotics were compared. RESULTS: The lpxM gene in AB AYE was successfully and fully knocked out. The absorbance value at OD600 for the lpxM knockout strain during the stable period was observed to be as low as 2.5, indicating a significant reduction in growth rate. Furthermore, the MIC of the knockout strain for imipenem decreased from 16 μg/mL to 1 μg/mL, and the MIC for ceftazidime decreased from 32 μg/mL to 16 μg/mL, enhancing antibiotic sensitivity. CONCLUSION: This study demonstrates that the deletion of the lpxM gene induces alterations in the growth and drug resistance of AB, providing a crucial foundation for further investigation into the mechanisms underlying LPS-mediated drug resistance and for the screening of effective auxiliary inhibitors targeting lpxM against MDR-AB.

特别声明

1、本页面内容包含部分的内容是基于公开信息的合理引用;引用内容仅为补充信息,不代表本站立场。

2、若认为本页面引用内容涉及侵权,请及时与本站联系,我们将第一时间处理。

3、其他媒体/个人如需使用本页面原创内容,需注明“来源:[生知库]”并获得授权;使用引用内容的,需自行联系原作者获得许可。

4、投稿及合作请联系:info@biocloudy.com。