Conjugation-Based Genome Engineering in Deinococcus radiodurans.

ABSTRACT: Deinococcus radiodurans has become an attractive microbial platform for the study of extremophile biology and industrial bioproduction. To improve the genomic manipulation and tractability of this species, the development of tools for whole genome engineering and design is necessary. Here, we report the development of a simple and robust conjugation-based DNA transfer method from E. coli to D. radiodurans, allowing for the introduction of stable, replicating plasmids expressing antibiotic resistance markers. Using this method with nonreplicating plasmids, we developed a protocol for creating sequential gene deletions in D. radiodurans by targeting restriction-modification genes. Importantly, we demonstrated a conjugation-based method for cloning the large (178 k

SUBMITTER: Brumwell SL 

PROVIDER: S-EPMC8939323 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature