A Multi-Layer-Controlled Strategy for Cloning and Expression of Toxin Genes in Escherichia coli.

ABSTRACT: Molecular cloning and controlled expression remain challenging when the target gene encodes a protein that is toxic to the host. We developed a set of multi-layer control systems to enable cloning of genes encoding proteins known to be highly toxic in Escherichia coli and other bacteria. The different multi-layer control systems combine a promoter-operator system on a transcriptional level with a riboswitch for translational control. Additionally, replicational control is ensured by using a strain that reduces the plasmid copy number. The use of weaker promoters (such as P_BAD or PfdeA) in combination with the effective theophylline riboswitch is essential for cloning genes that encode notoriously toxic proteins that directly target translation and transcript

SUBMITTER: Vandierendonck J 

PROVIDER: S-EPMC10467106 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature