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| - | <h1>Road to IGEM</h1> | + | <h1>Road to iGEM</h1> |
| - | <h2>Know the process we follow for our project. Make click on the phase you want to know.</h2> | + | <h2>Know the process we followed to design our project. Click on the phase you want to know.</h2> |
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Latest revision as of 03:49, 29 September 2011
Brainstorm
Road to iGEM
Know the process we followed to design our project. Click on the phase you want to know.
Salmonella is one of the principal microorganisms that cause food poisoning. It is a major concern for the food industry as it contaminates a wide range of food products quite easily. The impact on public health is considerable because Salmonella can be found in every country, so it can be considered a global public health issue. Developing countries have trouble keeping the sanitary conditions to prevent salmonellosis and typhoid fever that is why an effective method of detection is needed.
There are several methods for salmonella detection. Traditional methods such as the use of selective agar or standardized ISO methods can take up to 5 days. There are other rapid methods for detection and confirmation of Salmonella such as immunomagnetic separation, molecular methods like DNA hybridization and PCR assays, enzyme immunoassays (EIA), and enzyme-linked immunosorbent assays (ELISA). Some of these are claimed to give results within 48 hours.
The goal of this project is to develop a faster detection method using colorimetric techniques. Taking the base of the E. chromi project developed by the Cambridge 2009 iGEM team where there is no need to perform fluorescence microscopy.
A proposal is to develop a method to recombine Salmonella (if present) so that it produces a pigment visible to the human eye.
With the Systemic Lupus Erythematosus (SLE), the immunologic system will attack the healthy cells and tissues by mistake. This creates damage in the joints, skin, blood vessels and some organs. The SLE is one of the most common Lupus and it tends to affect a lot of parts of the body. The SLE is an autoimmune disease that causes the attack of the T cells to the healthy body cells, this causes the chronic inflammation.
The SLE can be mild o severe. Mild SLE can be controlled without many complications. Severe SLE can cause the death of the patient.
The cause of lupus has not been determined, but it's known that genes play an important role, although lupus is not determined just by them.
Our goal is to find an effective and specific treatment for systemic lupus erythematosus (SLE), minimizing secondary effects by attacking the reason of disease: over production of lymphocytes. This goal can be achieved by the altering of the immune dysregulation of SLE, interfering the pertinent apoptotic pathways. There are some genes, whose activation or deactivation, would nullify the problem of overproduced cells: Fas/Apo1/CD95.
The main objective of this proposal is to use designer bacteria to substitute the insulin injections to control glucose level in blood, by making the bacteria attach to the pancreas and then release the insulin when it is necessary.
Diabetes type 1 is a chronic disease that occurs when the pancreas does not produce enough insulin, which is a hormone that regulates the blood sugar levels. Nowadays, more than 220 million people in the world has diabetes, and in 2005 approximately 1.1 million people died from diabetes. It is believed that diabetes deaths will double between 2005 and 2030.
For that reasons, the project “Salmonella insullinae” would be a great option for the treatment of diabetes type I. The modeling of this project would consist in four phases: bacteria encapsulation, sensor, attachment, insulin production.
Predicted results:
- Obtain a designer bacteria to substitute the insulin injections to control glucose level in blood, by making the bacteria attach to the pancreas and release the insulin when it is necessary
- Doing in silica experiments and if it´s possible we could do some experiments in vivo with mice we can probe that the Salmonella insulinae is a theoretical real option for diabetics.
Dragline Silk
- Spider dragline silkis five times stronger by weight than steel, three times tougher than the top quality man-made fiber Kevlar.
- Highcommercialvalue
- Biomaterialsdevelopment
Goals
- To insert the gen MaSP1that codes for protein that is related with the synthesis of spider silk into an appropriate vector that could be BAC, plamid or cosmid. Make this technique to be standardized for further applications.
- To construct a system of biobricks that includes metabolic engineering.
Predicted results:
- Silk protein expression in flask cultivation.
- Production of silk proteins by HCDC.
- Purification of recombinant silk protein.
- Fiber spinning and mechanical testing.
- The insertion of the gen in a vector, which is capable to support it and include the aspect of metabolic engineering; construction on biobricks.
Presence of Aspergillus and its toxins represent a huge loss in the world production and in the food production income. (Abbas et al 2005)
Aflatoxins present important food safely problems in both developed and developing countries. Contamination is monitored in developed countries using enzyme‐linked immunusorbent assay (ELISA) ‐and high‐performance liquid chromatography (HPLC)‐based assays, both of which may be too expensive for routine use in many developing countries. There is a need for inexpensive alternative approaches to detect aflatoxins in lots of foods and feeds.
When ruminants are feed with food that has been contaminated with type B and G aflatoxins, these are metabolized end excreted through milk as type M1 and M2 aflatoxins. (Abbas, Hamed K. Aflotoxin and Food Safety. Boca Raton: Taylor & Francis Group, 2005)
The International agency for cancer research (IARC) has reported that aflatoxins from type B1 and M1 as possible human carcinogénics. (Food Tems and Constituents, Heterocyclic Aromatic Amines and Mycotoxins. Vol. 56, 1997) Both United States and European Union regulations on the MRL for M1 type aflatoxins of products that derive from milk establish a limit of 0.05mg of aflatoxins per Kg of sample.
Formal studies that involved the presence of M1 type aflatoxins in Mexico where done from 1990 to 1998. Until in 2008, when a research group from the Metropolitan Autonomous University at Xochimilco (UAM) quantified the presence of M1 type aflatoxins in raw, organic and ultra pasteurized milk that was produced in the mexican high plateau. Their results showed that the 59.1% of the tested milk contained dangerous levels of the M1 type aflatoxins. (Perez et al, n.d)
Chemotaxis is a conserved mechanism among bacteria that involves the molecular sensing of a variety of compounds important for cellular development. This mechanism enables the organism to detect a molecule of interest and move towards the concentration gradient by triggering several signaling pathways. The aim of this project is to design two biomolecular mechanisms that will enable the organism to detect and move towards a molecule of interest that would normally not be detected: Xylene. The first of such mechanisms consists of a xylene sensor which will trigger the expression of MCP protein Tsr-CCW, those proteins are the flagella movement regulators setting the Che family protein chain reaction that incites either smooth swimming or tumbling; The second mechanism will promote the expression of a reporter protein, allowing us to measure the concentration of xylene in the medium in a near real-time way.
Objectives
- Identify the presence of xylene in liquid media using colorimetric techniques.
- Construct a BioBrick to identify different concentrations of xylene using GFP.
- Construct a novel mechanism using the BioBrick model that promotes the movement of "E. coli" on the medium following the concentration gradient of a contaminant as xylene.
- Determine the appropriate way to insert the constructed plasmid into Escherichia coli RP8611 CGSC.
- Determine the social, economic and ecological implications of identifying xylene as industrial residue.
- Increased movement speed by the over expression of flagellum and the addition of ATC to the medium.
The ability of expressing extracellular pili-like appendages called nanowires that enable electron transportation (cellular respiration) in adverse (electron acceptor deficient) conditions, is a shared characteristic among several microorganisms. Shewanella oneidensis MR-1 is capable of producing such nanowires when grown in low concentrations of O2 and metal ions. The genes mshA, pilA, and gspG have been targeted as the main nanowire coding genes, while omcA, mtrC, and gspD confer conductivity to the nanowires as they code for the cytochromes needed for electron transport. This project intends to transfer these mechanisms into a more convenient model organism, Escherichia coli to create a synthetic strain capable of expressing electrically conductive nanowires. Preliminary experiments suggest that "E. coli" HB101 does not present nanowires, so it is a suitable organism for recombination.
Our goal is to create a recombinant "E. coli" strain, taking as a model "E. coli" HB101 and Shewanella oneidensis MR-1, capable of generating a difference of potential is the main objective of this project. In other words, we intend to integrate the electron conductivity mechanisms of S. oneidensis into "E. coli".
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