We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
An efficient system for markerless gene replacement applicable in a wide variety of enterobacterial species.
- Authors
White, A. P.; Allen-Vercoe, E.; Jones, B. W.; DeVinney, R.; Kay, W. W.; Surette, M. G.
- Abstract
We describe an improved allelic-exchange method for generating unmarked mutations and chromosomal DNA alterations in enterobacterial species. Initially developed for use in Salmonella enterica, we have refined the method in terms of time, simplicity, and efficiency. We have extended its use into related bacterial species that are more recalcitrant to genetic manipulations, including enterohemorrhagic and enteropathogenic Escherichia coli and Vibrio parahaemolyticus. Data from over 50 experiments are presented including gene inactivations, site-directed mutagenesis, and promoter exchanges. In each case, desired mutations were identified by polymerase chain reaction screening typically from as few as 10–20 colonies up to a maximum of 300 colonies. The method does not require antibiotic nor nutritional markers in target genes and works efficiently in wild-type strains, obviating the need for specialized hosts or genetic systems. The use is simple, requiring basic laboratory materials, and represents an alternative to existing methods for gene manipulation in the Enterobacteriaceae.
- Subjects
DNA replication; SALMONELLA enteritidis; ESCHERICHIA coli O157:H7; VIBRIO parahaemolyticus; POLYMERASE chain reaction; GENETIC mutation
- Publication
Canadian Journal of Microbiology, 2007, Vol 53, Issue 1, p56
- ISSN
0008-4166
- Publication type
Article
- DOI
10.1139/W06-102