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. 2009 Jul 13;37(17):e118. doi: 10.1093/nar/gkp561

Figure 2.

Figure 2.

The three-plasmid integron selection system. An example of the βDH10B host strain, which requires exogenous DAP for growth, carrying a pCC1FOS library clone containing gene cassettes is shown. The same strain also carries the pTRC99A::intI1 and R388ISceKT plasmids. IPTG-induced expression from pTRC99A::intI1 leads to integrase-mediated recombination between pCC1FOS plasmids containing attC sites and the attI1 site of the class 1 integron on the conjugative R388 plasmid. The recombinant plasmid is specifically selected by conjugation to the EPI300-T1R recipient and plating on media containing chloramphenicol but lacking DAP. The original class 1 integron of R388 is disrupted by the fusion. Small black arrows denote primers that can be used to sequence the gene cassettes of the recovered recombinant plasmids.