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. Author manuscript; available in PMC: 2013 Dec 5.
Published in final edited form as: J Am Chem Soc. 2012 Nov 20;134(48):19731–19738. doi: 10.1021/ja308049u

Figure 2.

Figure 2

Schematic procedures used for the deletion of a target chromosomal gene with the λ-Red recombination system42 (steps 1–3).40 In this work, the resistance cassette was removed from the new knock-out strain by FLP recombinase expressed from pCP20 vector43 (step 4),40 and a pACYC-based plasmid harboring tRNA genes (argU, ileY, and leuW) for the E. coli rare codons was subsequently incorporated into the resulting cells (step 5). FRT, FLP recombination target.43