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Team:Hawaii/Project
From 2008.igem.org
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Overall Project
Cyanobacteria are frequently studied for their ability to harness the power of photosynthesis to produce a wide variety of useful products including bio-fuels and -polymers. Such tasks are accomplished by these "little green factories" with a minimal input of salts, light, water, and carbon dioxide required for growth and carbon biomass accumulation. We aim to expand the availability of BioBrick vectors to cyanobacteria in order to “open source” the current BioBrick registry to a greater range of organisms.
We plan to engineer:
- 1) a mobilizable broad-host-range BioBrick vector that can be used to transfer genetic information between E. coli and Synechocystis sp. 6803, with the future possibility of transforming plants via Agrobacterium and other bacteria transformable by RSF1010 based plasmids;
- 2) a cassette for protein export from Synechocystis; and
- 3) the nitrate-inducible cyanobacterial nir promoter.
The functionality of the parts we engineer will be demonstrated by achieving inducible protein production and export of GFP construct introduced into Synechocystis using our novel BioBrick mobilizable shuttle vector.
Project Details
Part A: Mobilizable Broad-Host-Range Plasmid
- RSF1010 is a naturally occurring broad-host-range plasmid capable of conjugative transfer and stable replication due to the presence of mob genes with an associated origin of transfer (oriT) and rep genes with an associated origin of vegetative replication (oriV), respectively. We aim to compartmentalize a derivative of the RSF1010 plasmid, namely pRL1383a, into BioBricks. The resulting BioBricks can be inserted into a BioBrick base vector to create a plasmid that transfers genetic elements via conjugation. (read more...)
Part B: Cyanobacterial protein secretion system
- Photosynthetic cyanobacteria provide the opportunity for autotrophic production of practically any biomolecule. The ability to extract engineered biomolecules would make this bacterium a renewable, nearly self-sustaining "factory"- a potentially valuable tool in bioengineering. For Part B, we will create BioBricks encoding naturally occurring signal peptides that can be combined with a protein coding sequence in order to express the protein of interest extracellularly. (read more...)
Results
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
