Part improvement
Codon optimized LuxI
This year´s iGEM team Duesseldorf is contributing to the iGEM community by improving an existing part: (BBa_C0061) to a codon optimized version (BBa_K2587000). This part is originally from Aliivibrio fischeri and is a component of the quorum sensing system, a way of bacterial communication by cell population regulation. In this case luxI, the gene encoding 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 the modeling of 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 depends on different factors, such as synthetic transcription factors1.
In addition to that we added overhangs before and after the optimized coding sequence containing Type II S restriction sites for BsaI and BbsI. This second improvement allows to use this part directly for golden gate cloning, a popular cloning method, which further increases the modular cloning character.
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. (You can find a detailed explanation of our experiments here). Therefore, we codon optimized this sequence for the organism S. cerevisiae 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 optimizing a gene, that is not usually used in S. cerevisiae.
- Machens, Fabian, et al. "Synthetic Promoters and Transcription Factors for heterologous Protein expression in Saccharomyces cerevisiae." Frontiers in bioengineering and biotechnology 5 (2017): 63.