Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Ppublish 2 citations
PubMed: 36436701 DOI: 10.1016/j.mimet.2022.106627
Reporting off-target effects of recombinant engineering using the pORTMAGE system.
J Microbiol Methods 204:106627
Authors: Brittany R Sanders, Sydney E Townsend, Maria L Ford, Joseph L Graves, Misty D Thomas
Affiliations
- North Carolina Agricultural and Technical State University, USA.
- North Carolina Agricultural and Technical State University, USA.
Abstract
pORTMAGE recombineering is a simple technique for incorporation of novel point mutations into bacterial genomes that eliminates off-target effects. Here we inserted point mutations into the cusS gene from Escherichia coli, then, using Illumina sequencing, report genetic variants in all mutant strains. Several off-site mutations were found at high frequency. Low frequency mutations also show high heterogeneity. This means that it is essential for studies to report all off-target effects and acknowledge the effect that this may have on resultant phenotypes.
MeSH
Microbiological Techniques, Genes, Point Mutation