Abstract
Virulence traits and antibiotic resistance are frequently provided by genes located on plasmids. However, experimental verification of the functions of these genes is often lacking due to a lack of related experimental technology. In the present study, an integrated suicide vector was used to efficiently and specifically delete a bacterial endogenous plasmid in Cronobacter sakazakii. The pESA3 plasmid was removed from C. sakazakii BAA-894, and we confirmed that this plasmid contributes to the invasion and virulence of this strain. In addition, the pGW1 plasmid was expunged from C. sakazakii GZcsf-1, and we confirmed that this plasmid confers multidrug resistance. We further screened plasmid-curing agents and found that p-coumaric acid had a remarkable effect on the curing of pESA3 and pGW1 at sub-inhibitory concentrations. Our study investigated the contribution of endogenous plasmids pESA3 and pGW1 by constructing plasmid-cured strains using suicide vectors and suggested that p-coumaric acid can be a safe and effective plasmid-curing agent for C. sakazakii.
Highlights
Developed strategies for the treatment of systemic infections caused by pathogenic bacteria still rely on the application of various antibiotics
A strategy based on suicide vector pCVD442 was developed for plasmid curing in the current study
The plasmidspecific fragment of the suicide vector was taken from an endogenous plasmid (Figure 1), integration of the hybrid plasmid could be selected using the antibiotic corresponding to the resistance marker on the suicide vector
Summary
Developed strategies for the treatment of systemic infections caused by pathogenic bacteria still rely on the application of various antibiotics. The frequent and extensive utilization of antibiotics has dramatically increased the possibility of severe infections related to bacteria with multidrug resistance, posing a more aggressive threat to public health and necessitating the development of novel antimicrobials and bacterial vaccines. Related studies have unveiled the inseparable correlation between bacterial antibiotic resistance and plasmids, which could be verified by the frequent distribution of virulence determinants and antibiotic resistance genes in plasmids (Stratev and Odeyemi, 2016; Kopotsa et al, 2019). The abuse of antibiotics promoted the spread of antibiotic-resistant plasmids that have the potential to increase the occurrence of virulence or drug resistance genes, posing an increased threat to society (Fraise, 2002; Zhang et al, 2011; Mangat et al, 2017)
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