Generation of gene deletions and gene replacements in Escherichia coli O157:H7 using a temperature sensitive allelic exchange system

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Author: Emmerson, JR
Author: Roe, AJ
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Generation of gene deletions and gene replacements in Escherichia coli O157:H7 using a temperature sensitive allelic exchange system

James R. Emmerson¹, David L. Gally¹ and Andrew J. Roe¹^*

¹ Centre for Infectious Diseases, University of Edinburgh. The Chancellors Building, 49 Little France Crescent, EH16 4SB. United Kingdom.

* To whom correspondence should be addressed: Andrew J. Roe, Centre for Infectious Diseases, University of Edinburgh. The Chancellors Building, 49 Little France Crescent, EH16 4SB. United Kingdom. Email: aroe@vet.ed.ac.uk

Biol. Proced. Online 2006;8:153-162. doi:10.1251/bpo123
Submitted: July 05, 2006; Accepted: September 12, 2006; Published: September 27, 2006.

Indexing terms: Gene Deletion; Escherichia coli O157:H7.

Figure 2 Enlarged

Fig. 2:

(Adapted from Blomfield et al. 1991). Step 1: Plasmid integration in the intermediate strain is selected at the non permissive temperature for plasmid replication (42°C). Steps 2 and 3: Plasmid integrates are grown at 28°C in the absence of antibiotics to enrich for bacteria that have excised (Step 2) and later cured (Step 3) the integrated plasmid. Step 4: Growth on media containing sucrose selects for successful strain constructs.

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