Journal Article

Potentiation of curing by a broad-host-range self-transmissible vector for displacing resistance plasmids to tackle AMR

Lazdins, Alessandro, Haratianfar, Mona, Liu, Shuting, Haines, Anthony S., Kamruzzaman, Muhammad, Chamberlain, Melissa, Kreft, Jan-Ulrich, Miller, Claire E., Thomas, Christopher M., Maurya, Anand Prakash, Lloyd, Georgina S., Iredell, Jonathan, Webber, Mark. A., Stephens, Elton R.

2020

Overview

Plasmids are potent vehicles for spread of antibiotic resistance genes in bacterial populations and often persist in the absence of selection due to efficient maintenance mechanisms. We previously constructed non-conjugative high copy number plasmid vectors that efficiently displace stable plasmids from enteric bacteria in a laboratory context by blocking their replication and neutralising their addiction systems. Here we assess a low copy number broad-host-range self-transmissible IncP-1 plasmid as a vector for such curing cassettes to displace IncF and IncK plasmids. The wild type plasmid carrying the curing cassette displaces target plasmids poorly but derivatives with deletions near the IncP-1 replication origin that elevate copy number about two-fold are efficient. Verification of this in mini IncP-1 plasmids showed that elevated copy number was not sufficient and that the parB gene, korB, that is central to its partitioning and gene control system, also needs to be included. The resulting vector can displace target plasmids from a laboratory population without selection and demonstrated activity in a mouse model although spread is less efficient and requires additional selection pressure.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Addictions / Animals / Antibiotic resistance / Antibiotics / Antimicrobial agents / Bacteria / Bacterial Infections - drug therapy / Bacterial Infections - genetics / Bacterial Infections - microbiology / Binding sites / Biology and life sciences / Cassettes / Conjugation, Genetic - genetics / Control systems / Copy number / Curing / Disease Models, Animal / Displacement / DNA Copy Number Variations - genetics / DNA Primase - genetics / Drug resistance / Drug Resistance, Bacterial - genetics / Escherichia coli - genetics / Escherichia coli Proteins - genetics / Gastrointestinal Microbiome - drug effects / Gastrointestinal Microbiome - genetics / Host Specificity - genetics / Humans / Infections / Infectious diseases / Medical research / Medicine and Health Sciences / Mice / Microbiota / ParB gene / Physical Sciences / Plasmids / Plasmids - drug effects / Plasmids - genetics / Potentiation / Replication / Replication origins / Research and analysis methods / Vectors