Published: July 19, 2024

Despite the significant role of plasmids play in global spread antimicrobial resistance (AMR), current methods to study transfer antibiotic genes (ARGs) mainly rely on bacterial cultivation or sequencing techniques, leaving a knowledge gap understanding ARGs dissemination within natural microbial communities. To address this, new tools allowing real-time tracking are essential for comprehensive environmental risk assessments. Herein, we present culture-independent protocol examining horizontal across diverse populations. This method utilizes CRISPR-based editing fluorescently label wild-type AMR plasmids, facilitating their identification and sorting via Fluorescence-activated cell (FACS). As an illustrative example, detail step-by-step targeting gfp-tagged plasmid, followed by conjugation procedures FACS-based selection green fluorescent protein (GFP)-expressing transconjugants. fluorescence-based approach applied real-life populations can be uniquely deployed examine One Health factors such climate changes pollution.

Language: Английский