In Vitro Evolution to Increase the Titers of Difficult Bacteriophages: RAMP-UP Protocol.

ABSTRACT:

Background

Bacteriophages are becoming increasingly important in the race to find alternatives to antibiotics. Unfortunately, bacteriophages that might otherwise be useful are sometimes discarded due to low titers making them unsuitable for downstream applications.

Methods

Here, we present two distinct approaches used to experimentally evolve novel New Zealand Paenibacillus larvae bacteriophages. The first approach uses the traditional agar-overlay method, whereas the other was a 96-well plate liquid infection protocol that improved phage titers in as little as four days. We also used a mathematical model to probe the parameters and limits of the RAMP-UP approach to rapidly select mutants that improve bacteriophage titers.

Results

Both experimental approaches resulted

SUBMITTER: Kok DN 

PROVIDER: S-EPMC10282794 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature