Difference between revisions of "Team:Baltimore Bio-Crew/Safety"

 
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<header>
 
<header>
  
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                         <h3> About Our Project </h3>
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                         <h3>Safety</h3>
 
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<header>
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<h4>About our Lab</h4>
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</header>
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<article>
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Our lab is a community laboratory that is open to any skill level. The lab invites amateurs, professionals or just anyone interested in science. Due to this, our laboratory is a level one lab, in order to protect those working inside the lab, the general public and environment.
 +
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<article>
 
<article>
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<h4>Lab Safety</h4>
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Our goal for this project is to genetically engineer E. coli bacteria that can break down plastic. These bacteria could have many different applications, such as: degrading plastic waste from labs that cannot be recycled, being used in a filter to catch and degrade micro plastic fibers from laundry, and breaking down plastic in a marine environment into harmless molecules. We made a lot of progress last year, and this year we plan to build on that progress.
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</p>
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<p>
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<ol>
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While searching for solutions to the issue of plastic pollution in the Baltimore Inner Harbor, we found a paper by Yoshida et. al. describing a bacteria called Ideonella sakaiensis that was capable of degrading PET plastic into monomers. The bacteria used the enzyme PETase (chlorogenate esterase) to break down PET into MHET, and the enzyme MHETase (Lipase) to break down MHET into ethylene glycol and therephthalic acid. We decided to use the genes from this bacteria for our project.
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  <li>Laboratory Bootcamp
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</p>
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  <ul>
 +
    <li>Since we have different skill levels on our team, every member must participate in a yearly bootcamp.</li>
 +
    <li>The bootcamp allows team members to experience a laboratory setting and experiment with different commonly used lab tools, such as a pipette. The new students are supervised by the senior team members and by the laboratory’s directors.
 +
    <li>Also in the Bootcamp, team members learn the basics of biology and how to use safety equipment in the laboratory. Each member had to pass a test on where each piece of safety equipment was and what it was used for.
 +
    </li>
 +
  </ul>
 +
  </li>
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  <li>Community Lab Safety Rules
 +
  <ul>
 +
    <li>Wear proper lab equipment (gloves, goggles, and closed-toed shoes)</li>
 +
    <li>Disinfect the labspace (Sterilization, Disinfection, Antisepsis, and Cleaning)</li>
 +
    <li>Follow chemical safety procedures</li>
 +
    <li>Maintain a neat and well organized work environment</li>
 +
    <li>Use bleach to sterilize unused cultures</li>
 +
    <li>Keep food and drinks out of the lab</li>
 +
    <li>Dispose of chemical waste in the proper trash bin (chemical waste bins)</li>
 +
  </ul>
 +
  </li>
 +
</ol>
  
−
<p>
 
−
To avoid the safety risks of working with a relatively undocumented bacteria, we decided to take the plastic degradation genes from I. sakaiensis and put them into K12 E. coli bacteria. We chose E. coli because they are safe to work with and commonly used in the lab. Using the genetic sequence found in the paper, we designed the two plastic degrading enzymes so that they could be expressed in E. coli bacteria. We then had them synthesized and worked on putting these genes into E. coli.
 
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</p>
 
  
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<p>
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<h4>Project Safety</h4>
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By the end of last year’s competition, we had managed to insert the lipase gene into E.coli, but not the chlorogenate esterase gene. We confirmed that we had correctly inserted the lipase gene using colony PCR and gene sequencing, but we did not have the time to conduct additional assays, such as protein gels, to determine if the enzyme was being secreted from the bacteria. This year, we plan to redesign the chlorogenate esterase and lipase genes so that they contain the proper tags that will allow them to be detected, and a secretion sequence. After we insert both genes into E. coli cells, we will test them to make sure they can secrete the plastic degrading enzymes and degrade PET plastic.
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<ul>
−
+
  <li>Instead of using Ideonella sakaienisis in our laboratory, we used the K-12 strain of E.coli that was genetically modified to express the enzymes PETase (chlorogenate esterase) and MHETase (lipase). The K-12 strain of E.coli is a common bacterium used in laboratories while as Ideonella sakaienisis is fairly new in laboratories. Also, the enzymes we are expresses are not harmful and each team member knows the proper way to handle these enzymes in E.coli to prevent contamination.
 +
  </li>
 +
  <li>Since our bacteria may switch plasmids with other organisms in certain environments such as the Inner Harbor, the Baltimore BioCrew has decided to use alternative methods. The first would be containing out bacteria while also adding a kill switch. The kill switch would allow the bacteria to self-destruct if it detected the slightest malfunction, or escaped into the environment. The second option would be containing our bacteria in a bioreactor, where it would degrade any plastic added into the reactor.
 +
  </li>
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</ul>
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<h2>
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Sponsers
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Sponsors
 
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<h4>
 
<h4>
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</a>
 
</a>
  
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<a href="https://www.rwdfoundation.org/">
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                                      <a>
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   <img src="https://static.igem.org/mediawiki/2016/6/65/T--Baltimore_BioCrew--DeutschFoundation.png" alt="The Robert W. Deutsch Foundation" style="width:100px; height:100px;">
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   <img src="https://static.igem.org/mediawiki/2017/6/6c/T--Baltimore_Bio-Crew--fabian_kolker_small_icon.png" alt="Fabian Kolker Foundation" style="width:100px; height:100px;">
 
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</a>
  
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   <img src="https://static.igem.org/mediawiki/2016/1/1a/T--Baltimore_Biocrew--VWR_Foundation_LOGO.jpeg" alt="VWR Charitable Foundation" style="width:100px; height:100px;">
 
   <img src="https://static.igem.org/mediawiki/2016/1/1a/T--Baltimore_Biocrew--VWR_Foundation_LOGO.jpeg" alt="VWR Charitable Foundation" style="width:100px; height:100px;">
 
</a>
 
</a>
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<a href="http://vwrfoundation.org/">
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<a href="http://www.marylandrecyclingnetwork.org/">
 
   <img src="https://media.licdn.com/mpr/mpr/shrink_200_200/AAEAAQAAAAAAAAI8AAAAJDY0ZDg0ZjlkLWVlMTItNGI1Mi1iNWEwLWYzMDVlYWMwMTZhZg.png" alt="Maryland Recycling Network" style="width:100px; height:100px;">
 
   <img src="https://media.licdn.com/mpr/mpr/shrink_200_200/AAEAAQAAAAAAAAI8AAAAJDY0ZDg0ZjlkLWVlMTItNGI1Mi1iNWEwLWYzMDVlYWMwMTZhZg.png" alt="Maryland Recycling Network" style="width:100px; height:100px;">
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</a>
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<a href="https://www.rwdfoundation.org/">
 +
  <img src="https://static.igem.org/mediawiki/2016/6/65/T--Baltimore_BioCrew--DeutschFoundation.png" alt="The Robert W. Deutsch Foundation" style="width:100px; height:100px;">
 
</a>
 
</a>
  

Latest revision as of 19:31, 19 November 2017


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Safety

About our Lab

Our lab is a community laboratory that is open to any skill level. The lab invites amateurs, professionals or just anyone interested in science. Due to this, our laboratory is a level one lab, in order to protect those working inside the lab, the general public and environment.

Lab Safety

  1. Laboratory Bootcamp
    • Since we have different skill levels on our team, every member must participate in a yearly bootcamp.
    • The bootcamp allows team members to experience a laboratory setting and experiment with different commonly used lab tools, such as a pipette. The new students are supervised by the senior team members and by the laboratory’s directors.
    • Also in the Bootcamp, team members learn the basics of biology and how to use safety equipment in the laboratory. Each member had to pass a test on where each piece of safety equipment was and what it was used for.
  2. Community Lab Safety Rules
    • Wear proper lab equipment (gloves, goggles, and closed-toed shoes)
    • Disinfect the labspace (Sterilization, Disinfection, Antisepsis, and Cleaning)
    • Follow chemical safety procedures
    • Maintain a neat and well organized work environment
    • Use bleach to sterilize unused cultures
    • Keep food and drinks out of the lab
    • Dispose of chemical waste in the proper trash bin (chemical waste bins)

Project Safety

  • Instead of using Ideonella sakaienisis in our laboratory, we used the K-12 strain of E.coli that was genetically modified to express the enzymes PETase (chlorogenate esterase) and MHETase (lipase). The K-12 strain of E.coli is a common bacterium used in laboratories while as Ideonella sakaienisis is fairly new in laboratories. Also, the enzymes we are expresses are not harmful and each team member knows the proper way to handle these enzymes in E.coli to prevent contamination.
  • Since our bacteria may switch plasmids with other organisms in certain environments such as the Inner Harbor, the Baltimore BioCrew has decided to use alternative methods. The first would be containing out bacteria while also adding a kill switch. The kill switch would allow the bacteria to self-destruct if it detected the slightest malfunction, or escaped into the environment. The second option would be containing our bacteria in a bioreactor, where it would degrade any plastic added into the reactor.