Skip to main content
. Author manuscript; available in PMC: 2012 Dec 1.
Published in final edited form as: Methods. 2011 Sep 8;55(4):370–378. doi: 10.1016/j.ymeth.2011.08.019

Figure 1.

Figure 1

Schematic showing the deletion of a chromosomal gene with the λ-Red recombination system. Step 1 – Approximately 500 bp long upstream and downstream regions of the target gene were amplified from the E. coli chromosome by PCR. Note that each product has an overlap (black) with the resistance cassette. Step 2 – The linear double-stranded DNA was generated by PCR using the two DNA products from step 1 and the resistance cassette. Step 3 – The linear DNA was transformed into cells expressing λ-Red recombination system from pKD46. The new knock-out strain was selected used its resistance marker. Step 4 – If necessary, the resistance cassette was removed by FLP recombinase expressed from pCP20 vector[20].