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Team:WLC-Milwaukee
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This construct BBa_K875004 is designed for the expression of an already described engineered antinorovirus (NoV) monoclonal antibody (mAb 54.6) in fusion with LPP-OmpA. The antibody is expressed in a single chain fragment variable (scFv) format containing light (VL) and heavy (VH) variable domains separeted by a flexible peptide linker. It has already been reported that the scFv 54.6 binds a native recombinant NoV particles (VLPs) and inhibits VLP interaction with cells. LPP-OmpA functions as a leader sequence and an anchor to display the scFv ot the bacterial surface.The construct consistes of T5 Lac Operator (Bba_K875002), ribosomal binding site, LPP-OmpA-scFv 54.6 antinorovirus, Histidine tag (6HIS), Terminator (B0015). | This construct BBa_K875004 is designed for the expression of an already described engineered antinorovirus (NoV) monoclonal antibody (mAb 54.6) in fusion with LPP-OmpA. The antibody is expressed in a single chain fragment variable (scFv) format containing light (VL) and heavy (VH) variable domains separeted by a flexible peptide linker. It has already been reported that the scFv 54.6 binds a native recombinant NoV particles (VLPs) and inhibits VLP interaction with cells. LPP-OmpA functions as a leader sequence and an anchor to display the scFv ot the bacterial surface.The construct consistes of T5 Lac Operator (Bba_K875002), ribosomal binding site, LPP-OmpA-scFv 54.6 antinorovirus, Histidine tag (6HIS), Terminator (B0015). | ||
| + | <img src="https://static.igem.org/mediawiki/2013/2/2a/WLC-EcoliNissle1917.jpg" align="right" width=""> | ||
<h3>T5cumate repressed Tse2 mediated, antihorizontal transfer mechanism</h3> | <h3>T5cumate repressed Tse2 mediated, antihorizontal transfer mechanism</h3> | ||
This construct BBa_K1175003 is a modification of the part designed by iGEM2012 Team Trieste it has a T5cumate inducible promoter suppressed by the CymR gene integrated genome into the chassis E.coli Nissle 1917. This part will not be suppressed by CymR in the bacterial cells that have taken up the plasmid. | This construct BBa_K1175003 is a modification of the part designed by iGEM2012 Team Trieste it has a T5cumate inducible promoter suppressed by the CymR gene integrated genome into the chassis E.coli Nissle 1917. This part will not be suppressed by CymR in the bacterial cells that have taken up the plasmid. | ||
Revision as of 19:40, 27 September 2013
Our Project
This combinatory part is a self contained secretion system that combines a secretion and purification tag, a tripartite pump that is specific to the sec tag, a antinorovirus particle like antibody, Tse2 toxin mediated horizontal gene transfer mechanism.
Tripartite SecI pump
These are the three essential parts prtD, prtE, prtF, that make up the Type I secretion system from Erwinia chrysanthemi.This part is made of 3 genes: prtD, prtE, and prtF, that constitute a type I Erwinia chrysanthemi secretion system. The Pump is expressed from a strong constitutive promoter, BBa_k206000, and has the translational terminator BBa_B0014. In pSB1C3. Used to secrete proteins containing prtB C-terminal tag. The prtB C-terminal tag is built into the protein generator [BBa_K1175012]. Any protein of interest can be inserted into the protein generator and then secreted when used in conjunction with this secretion system.Secretion and purification tag
This part BBa_K1175012 A composite part formed between BBa_K206000 strong pBAD promoter, BBa_B0034 strong RBS, and BBa_K215001 a purification and secretion tag; a secretion system designed by Washington iGEM team 2009 BBa_K215001. This tag can be used to secrete proteins of interest by inserting them into a specific NheI cut site flanked by C terminus His tag and an N terminus Secretion tag. Both tags can be removed by a TEV protease. The secretion tag is also specific to the tripartite pump.Antinorovirus like particle antibody
This construct BBa_K875004 is designed for the expression of an already described engineered antinorovirus (NoV) monoclonal antibody (mAb 54.6) in fusion with LPP-OmpA. The antibody is expressed in a single chain fragment variable (scFv) format containing light (VL) and heavy (VH) variable domains separeted by a flexible peptide linker. It has already been reported that the scFv 54.6 binds a native recombinant NoV particles (VLPs) and inhibits VLP interaction with cells. LPP-OmpA functions as a leader sequence and an anchor to display the scFv ot the bacterial surface.The construct consistes of T5 Lac Operator (Bba_K875002), ribosomal binding site, LPP-OmpA-scFv 54.6 antinorovirus, Histidine tag (6HIS), Terminator (B0015).
T5cumate repressed Tse2 mediated, antihorizontal transfer mechanism
This construct BBa_K1175003 is a modification of the part designed by iGEM2012 Team Trieste it has a T5cumate inducible promoter suppressed by the CymR gene integrated genome into the chassis E.coli Nissle 1917. This part will not be suppressed by CymR in the bacterial cells that have taken up the plasmid.WHAT: We are repurposing cellulolytic genes to increase bioavailable energy by breaking down cellulose into glucose, which can be used for humanitarian efforts, economic mobility and Industrial application
WHY: Increase bioavailable fuel sources for human consumption, economic growth and increased reliance on local resources.
- Economic mobility: The cellulolytic probiotic strain that we are designing would be ideal for providing increased economic benefit and mobility to the underpriviledged populations that are the target individuals for this product. This would increase their economic powere by increasing the bioavailable resources that were once unavailable. This would do so by increasing nutrition in the cattle thus increasing the market value of the livestock, increasing the revenue available per capita. Also the increase in caloric availability could also apply to carbohydrate food sources thus allowing for an increased self sufficiency and, hopefully, the formation of a surplus that could enter into the world market. It is this entry into the world market that could facilitate a upward movement of under developed countries.
- Industrial application: The use of the cellulolytic E coli nissle 1917 would increase the availability of glucose for industrial application by opening a new avenue for using the renewable and abundant environmental cellulose. This could lead to an increased industrial self sufficiency by decreasing dependence on non renewable resources (i.e fossil fuels) or repurposed resources (i.e. corn and rice). In the case of fossil fuel, the world market depends on it due to its high demand but with the increased amount of bioavailable glucose there would be an increased availability of ethanol for use as a renewable fuel source. In addition with an increase in bioavailable glucose the world will be able to move away from high fructose corn syrup, which has been linked to an increased obesity rate in America, and towards high glucose syrup. This would mean a decreased reliance on corn allowing its use in other industrial or economic applications. All in all the use of a cellulolytic bacteria in industry would allow for a repurposing of what was once considered “trash” and instead turn it into a potential “treasure” using it as a resource, recycling what was once useless into something that can now be considered a reliable and renewable resource.
- Humanitarian efforts: Through the use of a known probiotic strain E. Coli Nissile 1917 the cellulolytic genes studied will be able to be used safely and effectively in human subjects. The effect of the cellulolytic probiotic strain would be to increase the caloric intake in the individuals who live as sustenance farmers, refugees, and in areas that do not provide enough agricultural means so as to provide sufficient calories for the population housed in the area. This increase in caloric intake would mean an increase in self sufficiency in third world countries and a decrease in economic strain for the supporting nations. The increase in self sufficiency would mean an increase in economic surplus within the country in question, eventually leading to an increase in the rate of development in these countries.
