Efficient Bacterial Genome Engineering throughout the Central Dogma Using the Dual-Selection Marker tetA^OPT.

ABSTRACT: Engineering of bacterial genomes is a fundamental craft in contemporary biotechnology. The ability to precisely edit chromosomes allows for the development of cells with specific phenotypes for metabolic engineering and for the creation of minimized genomes. Genetic tools are needed to select for cells that underwent editing, and dual-selection markers that enable both positive and negative selection are highly useful. Here, we present an optimized and easy-to-use version of the tetA dual-selection marker and demonstrate how this tetA^OPT can be used efficiently to engineer at different stages of the central dogma of molecular biology. On the DNA level, tetA^OPT can be used to create scarless knockouts across the Escherichia coli

SUBMITTER: Bayer CN 

PROVIDER: S-EPMC9594774 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature