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. Author manuscript; available in PMC: 2011 Jun 2.
Published in final edited form as: J Am Chem Soc. 2010 Jun 2;132(21):7519–7527. doi: 10.1021/ja102339q

Figure 4.

Figure 4

Schematic representation of knockout/knock-in strategy. (A) Excision of four structural gene copies via homologous recombination with a plasmid containing two areas of homology. (B) Knockout strain (exhibiting a 6 base pair scar from the initial knockout) is combined with a second plasmid baring a single area of homology and a single variant copy of the structural gene TclE. (C) Product variant strain is cultured and thiocillins isolated (pictured are comparative HPLC traces of isolates from the wild-type, the knockout, and a wild-type rescue).