Part improvement
Codon optimised LuxI
This year´s iGEM team Düsseldorf is contributing to the iGEM community by improving an existing part (BBa_C0061) to a codon optimised version (BBa_K2587000). This part is originally from Vibrio fischeri and is a component of the Quorum sensing system, a way of bacterial communication by cell population regulation. In this case LuxI, the acyl homoserine lactone synthase is described and improved. Most of the researchers working with the Quorum sensing module are interested in using this system in model prokaryotic organisms, such as Escherichia coli. Nonetheless more and more scientists are trying to implement similar systems in eukaryotic organisms as well, which should not be neglected. In fact, designing synthetic gene regulatory circuits for example in Saccharomyces cerevisiae depend on different factors, such as synthetic transcription factors1).
Experimental design
Part of our project is the design of a synthetic promoter that is able to induce expression of a reporter gene, which is activated upon synthesis of the Quorum sensing molecule acyl homoserine lactone by the respective synthase LuxI. (Hier link to QS Experimental design Seite). Therefore, we codon optimised this sequence for the organism S.cerervisiae and implemented this in our construct. Thus, we did not only apply this sequence into our project, but also improved a previous part by codon optimising a gene, usually not used for S.cerevisiae. In addition to that, our sequence contains Type II S restriction sites, which can be used for Golden Gate cloning, a trending cloning method.
