Measuring differential fitness costs and interactions between genetic cassettes using fluorescent spectrophotometry.

ABSTRACT: In this article, we present a method for designing, executing, and analyzing data from a microbial competition experiment. We use fluorescent reporters to label different competing strains and resolve individual growth curves using a fluorescent spectrophotometer. Our comprehensive data analysis pipeline integrates multiple experiments to simultaneously infer sources of variation, extract selection coefficients, and estimate the genetic contributions to fitness for various synthetic genetic cassettes (SGCs). To demonstrate the method, we employ a synthetic biological system based on Escherichia coli. Strains carry 1 of 10 different plasmids and one of three genomically integrated fluorescent markers. All strains are co-cultured to obtain real-time measurements of optical density (to

SUBMITTER: Kehila D 

PROVIDER: S-EPMC10880626 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature