Team:Cornell/project/drylab/functionalreq

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<h1 style="margin-top: 0px;">Header 1</h1>
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<h1>Functional Requirements</h1>
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Our system needed to allow diffusion of heavy metal ions across a filter boundary without letting the genetically engineered cells out into the environment.  To satisfy this requirement, we bought a hollow fiber reactor with a MWCO of 5kd @ 50%, effectively isolating the E.coli cells(.7-1.4micrometers)<sup>[1]</sup> from the outlet, while allowing diffusion of smaller molecules such as ions across the membrane of the fibers.
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<b>Filter Out Heavy Metals</b>
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Our system needed to allow diffusion of heavy metal ions across a filter boundary without letting the genetically engineered cells out into the environment.  To satisfy this requirement, we bought a hollow fiber reactor with a molecular weight cut-off (MWCO) of 20 kd @ 50%, effectively isolating the E.coli cells (0.7-1.4 micrometers) from the outlet,<sup>[1]</sup> while allowing diffusion of smaller molecules such as ions across the membrane of the fibers.
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Additionally, water needed to have an initial flow rate that would allow it to pass through the filter with the cells completely while still allowing time for the diffusion of any heavy metals across the filter membrane to be taken up by the cells. This was accomplished by placing a pump between the reservoir and the hollow fiber reactor.
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<b>Isolated System</b>
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Additional components were added to improve the system, including a carbon water filter before the hollow fiber reactor in order to filter out large particles, which would clog the hollow fiber reactor, and to purify the water of certain compounds which would harm the cells. An additional requirement for the pump in the system was that it had to move water through the large filter first.  Also, a battery was required to power the pump. Since this system was modeled to work outside for extended periods of time attached to a factory outlet pipe, a solar panel was used to power a long lasting battery. Finally, a bucket functioned as the reservoir to collect the water from the pipe.
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The entire filter system is housed in a watertight box to prevent contaminants from entering. To filter out debris that could enter via the collection bucket, a carbon filter is placed in the system before the fiber reactor. This ensures that only water with microscopic contaminants enters the fiber reactor. Thus, the system is as isolated from the outside environment as possible.
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The system was scaled down slightly for our purposes, since volume of water was not a concern, so only one hollow fiber reactor to house the cells was necessary.
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<br><br>
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<b>Effective Flow Rates</b>
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A carbon water filter was added before the hollow fiber reactor in order to filter out large particles and debris that would clog the hollow fiber reactor and to purify the water of certain compounds which would harm the cells. Entering water needed to have an initial flow rate that would allow it to pass through both filters while still allowing time for heavy metal diffusion across the fiber reactor membrane and uptake by the cells. The pump was placed before the carbon fiber in order to move it effectively through the carbon fiber and still have a slow flow rate through the fiber reactor.  
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<li>
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<b>Saturation Detection</b>
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A subsystem of reporter constructs was set up inside the box in order to detect when the metallothionein in the fiber reactor is saturated with heavy metals and the cells need replacing. The constructs consisted of a chromoprotein, amilCP, downstream from a specific heavy metal inducible promoter. When the metallothioneins became concentrated, the cells turn visibly blue, indicating that the cells need to be replaced.
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<b>Maintenance</b>
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Our system is powered by a high capacity car battery that is recharged by a compact solar panel. Since our flow requirements are not high, the system can last for up to 2 weeks without any sunlight at all. With several hours of sunlight a day, the system can retain charge for months at a time. Combined with the tough, weatherproof design, this allows our system to purify water fully autonomously for very long periods.  
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Far far away, behind the word mountains, far from the countries Vokalia and Consonantia, there live the blind texts. Separated they live in Bookmarksgrove right at the coast of the Semantics, a large language ocean. A small river named Duden flows by their place and supplies it with the necessary regelialia.
 
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Far far away, behind the word mountains, far from the countries Vokalia and Consonantia, there live the blind texts. Separated they live in Bookmarksgrove right at the coast of the Semantics, a large language ocean. A small river named Duden flows by their place and supplies it with the necessary regelialia.
 
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But I must explain to you how all this mistaken idea of denouncing pleasure and praising pain was born and I will give you a complete account of the system, and expound the actual teachings of the great explorer of the truth, the master-builder of human happiness.
 
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The quick, brown fox jumps over a lazy dog. DJs flock by when MTV ax quiz prog. Junk MTV quiz graced by fox whelps. Bawds jog, flick quartz, vex nymphs. Waltz, bad nymph, for quick jigs vex! Fox nymphs grab quick-jived waltz. Brick quiz whangs jumpy veldt fox. Bright vixens jump; dozy fowl quack.
 
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No one rejects, dislikes, or avoids pleasure itself, because it is pleasure, but because those who do not know how to pursue pleasure rationally encounter consequences that are extremely painful. Nor again is there anyone who loves or pursues or desires to obtain pain of itself, because it is pain, but because occasionally circumstances occur in which toil and pain can procure him some great pleasure. To take a trivial example, which of us ever undertakes laborious physical exercise, except to obtain some advantage from it? But who has any right to find fault with a man who chooses to enjoy a pleasure that has no annoying consequences, or one who avoids a pain that produces no resultant pleasure? On the other hand, we denounce with righteous indignation and dislike men who are so beguiled and demoralized by the charms of pleasure of the moment, so blinded by desire, that they cannot foresee.
 
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<h1>Header 3</h1>
 
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The quick, brown fox jumps over a lazy dog. DJs flock by when MTV ax quiz prog. Junk MTV quiz graced by fox whelps. Bawds jog, flick quartz, ex nymphs. Waltz, bad nymph, for quick jigs vex! Fox nymphs grab quick-jived waltz. Brick quiz whangs jumpy veldt fox. Bright vixens jump; dozy fowl quack.
 
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<h1>References</h1>
<h1>References</h1>
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<li>Nelson DE, Young KD. Penicillin binding protein 5 affects cell diameter, contour, and morphology of Escherichia coli. J Bacteriol. 2000 Mar182(6):1714-21 p.1719</li>
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<li>Nelson DE, Young KD. Penicillin binding protein 5 affects cell diameter, contour, and morphology of Escherichia coli. <i>J Bacteriol.</i> 2000 Mar182(6):1714-21 p.1719.</li>
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<li>Ref 3</li>
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Latest revision as of 03:49, 18 October 2014

Cornell iGEM

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Dry Lab

Functional Requirements

  1. Filter Out Heavy Metals Our system needed to allow diffusion of heavy metal ions across a filter boundary without letting the genetically engineered cells out into the environment. To satisfy this requirement, we bought a hollow fiber reactor with a molecular weight cut-off (MWCO) of 20 kd @ 50%, effectively isolating the E.coli cells (0.7-1.4 micrometers) from the outlet,[1] while allowing diffusion of smaller molecules such as ions across the membrane of the fibers.
  2. Isolated System The entire filter system is housed in a watertight box to prevent contaminants from entering. To filter out debris that could enter via the collection bucket, a carbon filter is placed in the system before the fiber reactor. This ensures that only water with microscopic contaminants enters the fiber reactor. Thus, the system is as isolated from the outside environment as possible.
  3. Effective Flow Rates A carbon water filter was added before the hollow fiber reactor in order to filter out large particles and debris that would clog the hollow fiber reactor and to purify the water of certain compounds which would harm the cells. Entering water needed to have an initial flow rate that would allow it to pass through both filters while still allowing time for heavy metal diffusion across the fiber reactor membrane and uptake by the cells. The pump was placed before the carbon fiber in order to move it effectively through the carbon fiber and still have a slow flow rate through the fiber reactor.
  4. Saturation Detection A subsystem of reporter constructs was set up inside the box in order to detect when the metallothionein in the fiber reactor is saturated with heavy metals and the cells need replacing. The constructs consisted of a chromoprotein, amilCP, downstream from a specific heavy metal inducible promoter. When the metallothioneins became concentrated, the cells turn visibly blue, indicating that the cells need to be replaced.
  5. Maintenance Our system is powered by a high capacity car battery that is recharged by a compact solar panel. Since our flow requirements are not high, the system can last for up to 2 weeks without any sunlight at all. With several hours of sunlight a day, the system can retain charge for months at a time. Combined with the tough, weatherproof design, this allows our system to purify water fully autonomously for very long periods.

References


  1. Nelson DE, Young KD. Penicillin binding protein 5 affects cell diameter, contour, and morphology of Escherichia coli. J Bacteriol. 2000 Mar182(6):1714-21 p.1719.