TRNA-Mediated Plasmid Stabilization for Antibiotic-Free Applications in <i>Escherichia coli</i>.

ABSTRACT: Plasmids are essential tools in molecular biology and biotechnology. In research laboratories, it is common to use antibiotic selection markers to ensure that plasmids are stably maintained in a cellular population. However, the use of antibiotics poses a significant challenge in the industrial scale-up process due to the high cost and the risk of spreading resistance. Therefore, methods for antibiotic-free plasmid maintenance are in high demand. Here, we present an essential gene-based plasmid selection strategy utilizing the Escherichia coli tryptophan tRNA (trpT) gene. We developed a workflow using a base strain with a trpT deletion and a temperature-sensitive trpT-expressing plasmid to circumvent the need for remaking chromosomal trpT deletions for ev

SUBMITTER: Sepulchro AGV 

PROVIDER: S-EPMC12814533 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature