The first host organism we will be working on is the cyanobacterium Synechocystis sp. strain PCC 6803sp. PCC 6803. Cyanobacteria constitute a unique class of microorganisms that are able to fix and metabolize carbon dioxide using the energy derived from sunlight. Cyanobacteria also present some of the highest solar energy conversion efficiencies found in nature (Dismukes et al., 2008). As cyanobacteria require nutrients like phosphorus and nitrogen for growth, they can also theoretically be used to sequester these pollutants from both industrial and agricultural streams, thereby further reducing production costs and environmental impact. Synechocystis exhibits robust and predictable growth in laboratory settings and the effect of different parameters including salt concentration, pH, temperature, UV light and CO2 level on its growth has been investigated (Lopo et al., 2012). Despite being one of the most extensively studied cyanobacterial species, the use of Synechocystis as a synthetic biology chassis awaits further investigation.
Competent cells were obtained following the iGEM protocol Help:Protocols/Competent Cells
E. coli DH5α was transformed using the protocol Help:Protocols/Transformation
The Fluorescence Intensity was measured using the standardized protocol from iGEM Plate_Reader_Protocol_Update . The Plate Reader (Fluostar omega, BMG LABTECH) was calibrated using the solutions included in the Interlab Measurement Kit.
Fluorescence Intensity was measured using the Flow Cytometer (Attune NxT, Thermo Fisher Scientific) in cells grown in LB following guidelines from iGEM. The Flow Cytometer was calibrated using Sphero®Rainbow Calibration Particles (BD Bioscience), 8 peaks, calibrated for MEFL (Molecules of Equivalent Fluorescein). Four drops of calibration particles were dissolved in sheath fluid (1ml). Samples were prepared for measurement in the Flow Cytometer washing the culture media in filtered 1X PBS. Cells cultures were diluted 1:100, adding PSB in a 96-well microtiter plate (Thermofisher Scientific). The instrument was configured with a channel for GFP measurement with 488 nm laser and 530/30 filter
We are the University of Edinburgh Overgraduate iGEM Team, competing in the new application track in iGEM 2016. read more
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Email: edigemmsc@ed.ac.uk