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- Title
FRUIT, a scar-free system for targeted chromosomal mutagenesis, epitope tagging, and promoter replacement in Escherichia coli and Salmonella enterica.
- Authors
Stringer, Anne M; Singh, Navjot; Yermakova, Anastasiya; Petrone, Brianna L; Amarasinghe, Jayaleka J; Reyes-Diaz, Lucia; Mantis, Nicholas J; Wade, Joseph T
- Abstract
Recombineering is a widely-used approach to delete genes, introduce insertions and point mutations, and introduce epitope tags into bacterial chromosomes. Many recombineering methods have been described, for a wide range of bacterial species. These methods are often limited by (i) low efficiency, and/or (ii) introduction of "scar" DNA into the chromosome. Here, we describe a rapid, efficient, PCR-based recombineering method, FRUIT, that can be used to introduce scar-free point mutations, deletions, epitope tags, and promoters into the genomes of enteric bacteria. The efficiency of FRUIT is far higher than that of the most widely-used recombineering method for Escherichia coli. We have used FRUIT to introduce point mutations and epitope tags into the chromosomes of E. coli K-12, Enterotoxigenic E. coli, and Salmonella enterica. We have also used FRUIT to introduce constitutive and inducible promoters into the chromosome of E. coli K-12. Thus, FRUIT is a versatile, efficient recombineering approach that can be applied in multiple species of enteric bacteria.
- Publication
PloS one, 2012, Vol 7, Issue 9, pe44841
- ISSN
1932-6203
- Publication type
Journal Article
- DOI
10.1371/journal.pone.0044841