Team:Hawaii/Project

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== Overall Project ==
== Overall Project ==
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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.
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'''A BioBrick toolkit for cyanobacteria'''
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We plan to engineer:<br>
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: 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; <br>
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: 2) a cassette for protein export from ''Synechocystis''; and
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: 3) the nitrate-inducible cyanobacterial nir promoter.
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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.
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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.
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We designed:
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1) mobilizable broad-host range BioBrick vectors derived from RSF1010,
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2) a cassette for protein secretion from ''Synechocystis'' sp. PCC 6803, and  
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3) a nitrate-inducible cyanobacterial promoter BioBrick.
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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==
== Project Details==

Revision as of 21:31, 21 October 2008

Projects Events Resources
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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.

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...)

Results

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