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Team:Hawaii/Project
From 2008.igem.org
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Overall Project
A BioBrick toolkit for cyanobacteria
- We aim to extend the current BioBrick registry to a greater range of organisms, including cyanobacteria. Cyanobacteria are studied for their ability to produce useful compounds, including biofuels and biopolymers. These "little green factories" require only salts, light, water, and carbon dioxide for photoautotrophic growth. A cyanobacterial "toolkit" would enhance our ability to utilize this system.
- We designed:
- 1) mobilizable broad-host range BioBrick vectors derived from RSF1010,
- 2) a cassette for protein secretion from Synechocystis sp. PCC 6803, and
- 3) a nitrate-inducible cyanobacterial promoter BioBrick.
- 1) mobilizable broad-host range BioBrick vectors derived from RSF1010,
- Our toolkit was designed for conjugative gene transfer from Escherichia coli to Synechocystis to achieve the controlled production and recovery or bioproducts, demonstrable by induced secretion of green fluorescent protein. Though our parts were targeted for work in cyanobacteria, they may be compatible with other Gram-negative systems including Agrobacterium, which is capable of plant transformation.
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...)
Materials, Methods, and Results
Part A: Mobilizable Broad-Host-Range Plasmid
Part B: Expression system for Synechocystis sp. PCC 6803
Implications and future plans
