Team:Copenhagen/Project

Project

Project description

Conventional single celled model organisms, such as E.coli, are widely used in synthetic biology as reliable and well-characterized systems. However a carbon rich medium has to be supplied which is expensive in both economic and environmental terms. Using cyanobacteria capable of performing photosynthesis and removing waste CO2 bypasses this problem and has the potential to be an important organism for future biotechnological research. We wish to genetically engineer cyanobacteria to produce biologically generated light, so-called bioluminescence. This not only has direct implications for development of sustainable and economic light sources, but also as a general proof-of-principle of the suitability of cyanobacteria for biotechnological research and production.

General Experimental Design

Initially the project sets with identification of native promoters from cyanobacteria responding to the day-night-cycle. Those will be spliced into an iGEM-biobrick together with the luxCDABE cassette containing the essential luciferase enzyme (and auxillary enzymes necessary for its function). The process is carried out in E.coli, and afterwards the complete plasmid is transferred to cyanobacteria (strain PCC6803). The endogenous transcription factors present in cyanobacteria should appropriately active transcription from the genetically spliced promoter thus driving transcription only during nighttime. In conjunction we will as a proof-of-principle experiment use the already existing plasmids pDawn and pDusk and BioBricks from the iGEM Registry, to, in a similar manner, create a luciferase containing plasmid, but with an artificial promoter system responding directly to light-dark conditions. It serves as a parallel control experiment. Both of these systems will subsequently be thoroughly analyzed to determine important characteristics of the system including kinetics and efficiency of the expression levels. To achieve this quantification we will collaborate with fellow Mathematics and Physics students at University Copenhagen. In this our date can be compared to simulations. As an example it be possible to put CO2 consumption by the cyanobacteria in relation to the emitted bioluminescence thus describing the degree of sustainability or ‘greenness’ of the system.