A New Suite of Allelic-Exchange Vectors for the Scarless Modification of Proteobacterial Genomes.

ABSTRACT: Despite the advent of new techniques for genetic engineering of bacteria, allelic exchange through homologous recombination remains an important tool for genetic analysis. Currently, sacB-based vector systems are often used for allelic exchange, but counterselection escape, which prevents isolation of cells with the desired mutation, occasionally limits their utility. To circumvent this, we engineered a series of "pTOX" allelic-exchange vectors. Each plasmid encodes one of a set of inducible toxins, chosen for their potential utility in a wide range of medically important proteobacteria. A codon-optimized rhaS transcriptional activator with a strong synthetic ribosome-binding site enables tight toxin induction even in organisms lacking an endogenous rhamnose regulon. Expressi

SUBMITTER: Lazarus JE 

PROVIDER: S-EPMC6677854 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature