Lambda Red-mediated recombinogenic engineering of enterohemorrhagic and enteropathogenic E. coli.

ABSTRACT:

Background

The lambda Red recombineering technology has been used extensively in Escherichia coli and Salmonella typhimurium for easy PCR-mediated generation of deletion mutants, but less so in pathogenic species of E. coli such as EHEC and EPEC. Our early experiments with the use of lambda Red in EHEC and EPEC have led to sporadic results, leading to the present study to identify factors that might improve the efficiency of Red recombineering in these pathogenic strains of E. coli.

Results

In this report, we have identified conditions that optimize the use of lambda Red for recombineering in EHEC and EPEC. Using plasmids that contain a Ptac-red-gam operon and a temperature-sensitive origin of replication, we have generated multiple mutations (both marked and unmarked) in kn

SUBMITTER: Murphy KC 

PROVIDER: S-EPMC317293 | biostudies-literature | 2003 Dec

REPOSITORIES: biostudies-literature