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Validation of Chromosome Recombination with λ-Red Technique

In order to test if the basic genome-editing part---λ-Red system works, we designed several different knock-in site pairs on E.coli chromosome and utilize antibiotic resistance gene to target them.

Firstly, from a large number of paper collection, we summarized aλ-Red protocol which tells that recombination bacteria cell preparation requires L-arabinose induction, and L-arabinose appears to inhibit cell growth due to cytotoxicity ofλ-Red proteins. Therefore, as to learn about cell growth circumstances under L-arabinose induction, and test ifλ-Red proteins get expressed on some level, we made a growth curve of gradient arabinose induced cell growth as follows. Through the curve, we can easily find that with the increase of arabinose induced concentration, cell grows slower and slower, implying the inhibition effect got strengthened.

Figure 1.The growth curves under L-arabinose gradients induction. Inhibition effect got strengthened with the increase of arabinose induced concentration.

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