Abstract
AbstractDNA nucleases TnpB and IscB were regarded as new antibacterial strategy to combat the drug-resistant bacteria represented byEscherichia colidue to its specificity in targeting DNA and smallest size, but the genome-editing of TnpB/IscB inE. coliremains unclear. This study characterized the genome-editing of TnpB/IscB inE. colistrains. First, the toxicity and cleavage results showed TnpB only worked inE. coliMG1655, while IscB and enIscB could perform in ATCC9637 and BL21(DE3). Next, TnpB-based genome-editing tool was established in MG1655, while IscB/enIscB achieved in ATCC9637/BL21(DE3). The copy number of TnpB/IscB/enIscB were changed to explore the impact of editing efficiency. Moreover, the editing plasmids were successfully cured. Finally, the escaping mechanism ofE. coliunder editing of TnpB/IscB was revealed. Overall, this study successfully applied TnpB/IscB/enIscB to genome-editing inE. coli, which will broaden genetic manipulation toolbox inE. coliand facilitate the development of new antimicrobial drugs.
Publisher
Cold Spring Harbor Laboratory
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