Creation of a promoter library

Site directed saturation mutagenesis PCR of the BBa_R0062 PluxR

Figure 1: Promoter mutation sites The wild type promoter from the biobrick BBa_R0062 was mutated using site directed saturation mutagenesis. The library shows the targeted sites for site directed saturation mutagenesis. The sequences for the library, the BBa_R0062, the BBa_K1216007 and the wild type luxR promoter from literature(2) are given. All assays were carried out in duplicates, results are presented as mean ± standard deviation.

We did site directed saturation mutagenesis of specific sites of the lux box according to the results taken from literature (2) (see Figure 2).
We mutated the promoter directly in the BBa_J09855 construct and cloned it into a eGFP gene as a reporter to be able to screen for differences in the dose response curves. In the end we isolated one mutated promoter which shows a shift in sensitivity (see section below)

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