Subject Code : BTEC8002
Country : Australia
Assignment Task:

Experiment design flow overview and experimental designs for ectoine expression in Synechococcus elongatus PCC 7942 using the ectoine gene cassette of the extromophilic bacterium Halomonas elongate 

Background brief on global challenges: The world is facing a carbon and freshwater constrained economy, which is exacerbated by finite resources (fossil fuel & land). The global population will be exceeding 9 billion people by 2050, which will place additional demand on resources with increases of 70, 50, and 30% being predicted for food, fuel, and water, respectively by 2050. Agricultural production to meet the predicted 70% increase is challenged by scarce freshwater availability and greenhouse gas emission induced climatic instability, as well as diminishing arable land. Algae, including the photosynthetic cyanobacteria to which Synechococcus elongatus belongs, have been heralded to be the potential saviours, as the fix atmospheric carbon dioxide (a greenhouse gas), converting it to organic carbon compounds, such as protein, carbohydrates and lipids, and they can be grown on non-arable land in non-potable water. 

Problem statement for commercial production of microalgae for greenhouse gas emission abatement and animal protein replacement in human diets: Climatic instability will lead to unpredictable rainfall patterns (i.e. draughts and floods), as well as temperature (generally the world is getting hotter). While algae can be potential saviours, they generally do not grow well at temperatures exceeding 300C. While some can withstand increased water salinities, some cannot. Hence commercial production of microalgae for greenhouse gas emission abatement and a source of protein to replace agricultural-derived protein (primarily animal protein) is not going to be easily achieved. To accomplish this goal, temperature and salinity resilient microalgal strains are required. 

Background brief on a potential pathway to obtain temperature and salinity resilient microalgae: Extremophilic bacteria, such as Halomonas elongate can survive under extreme heat & salinity, but they are not a suitable source of protein for human diets and they do not fix carbon dioxide. Many extremophilic bacteria accumulate an osmolyte called ectoine. Ectoine is a hydrophilic amino acid that infers tolerance to high salinity and temperature. Ectoine is a strong osmo-protectant (against saline-induced osmotic stress), and also protects against UV radiation and dehydration, which is essential to avoid protein misfolding, protection of DNA, RNA, and maintenance of membrane fluidity. Ectoine has also been shown to protect against thermal stress and maintenance of photosynthetic rates. 

Tasks:
1) Design and construct a suitable vector for transforming S. elongatus with the ectABC gene cassette from H. elongata
2) To produce a detailed experimental design flow chart that outlines succinctly all steps required.
3) Produce separate detailed specific experimental designs for all required processes.
4) Based on the above, write a materials and methods section, referring back to the produced diagrams as figures.

 

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  • Posted on : June 18th, 2019
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