A new recombineering system for precise genome-editing in Shewanella oneidensis strain MR-1 using single-stranded oligonucleotides.

ABSTRACT: Shewanella oneidensis MR-1 is an invaluable host for the discovery and engineering of pathways important for bioremediation of toxic and radioactive metals and understanding extracellular electron transfer. However, genetic manipulation is challenging due to the lack of genetic tools. Previously, the only reliable method used for introducing DNA into Shewanella spp. at high efficiency was bacterial conjugation, enabling transposon mutagenesis and targeted knockouts using suicide vectors for gene disruptions. Here, we describe development of a robust and simple electroporation method in S. oneidensis that allows an efficiency of ~4.0 x 10⁶ transformants/µg DNA. High transformation efficiency is maintained when cells are frozen for long term storage. In addition, we report a new p

SUBMITTER: Corts AD 

PROVIDER: S-EPMC6328582 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature