Difference between revisions of "Team:UCSC/Safety"

 
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</style>
 
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<div id="page">
 
<div id="page">
 
<center>
 
<center>
 
<br>
 
<br>
  
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<img class="titleimg" src="https://static.igem.org/mediawiki/2017/3/37/Gloves.png">
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<img class="titleimg" src="https://static.igem.org/mediawiki/2017/3/37/Gloves.png" alt="http://freevector.co/vector-icons/fashion/protection-gloves.html">
 +
 
 +
<h1>SAFETY</h1>
  
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<h1>SAFETY</h1><br>
 
 
<br>
 
<br>
 
<br>
 
<br>
  
 
<div class="text-container">
 
<div class="text-container">
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   <p><span class="firstword">When</span> choosing an organism to help us produce acetaminophen and vitamin B-12 we sought out an FDA approved microbe; however, we resorted to using a non-toxic cyanobacteria as a model for spirulina. Spirulina is generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.
+
   <p><span class="firstword">When</span> choosing an organism to help us produce acetaminophen and vitamin B<sub>12</sub> we sought out an FDA-approved microbe; however, we resorted to using a non-toxic cyanobacteria, <i>Synechococcus elongatus</i> strain PCC 7942, as a model for <i>Arthrospira platensis</i>. <i>A. platensis</i> is a generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.</p>
−
  </p>
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   <div class="container">
 
   <div class="container">
 
       <div class="row">
 
       <div class="row">
 +
          <h2><b>L A B</b>  SAFETY</h2>
 +
          <br>
 
           <div class="col-md-1"></div>
 
           <div class="col-md-1"></div>
 
           <div class="col-md-5">
 
           <div class="col-md-5">
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            <h2>LAB SAFETY</h2>
 
 
             <img src="https://static.igem.org/mediawiki/2017/d/d2/Safety-evan.png" style="width: 95%;">
 
             <img src="https://static.igem.org/mediawiki/2017/d/d2/Safety-evan.png" style="width: 95%;">
 
           </div>
 
           </div>
 
       <!-- </div> -->
 
       <!-- </div> -->
 
           <div class="col-md-5">
 
           <div class="col-md-5">
−
           <p>Proper PPE for BSL-1 organisms are worn in our laboratory. Within the lab space, every person is expected to be wearing gloves, long pants, close-toed shoes, a lab coat, and protective goggles. All personnel also must be accompanied with one other person so there is no one alone in lab.
+
            <br><br><br><br>
 +
           <p>Proper PPE for BSL-1 organisms are worn in our laboratory. Within the lab space, every person is expected to be wearing gloves, long pants, close-toed shoes, a lab coat, and protective goggles. All personnel also must be accompanied with one other person so that there is no one alone in lab.
 
           </p>
 
           </p>
 
           </div>
 
           </div>
 
       </div>
 
       </div>
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   </div><br><br><br><br><br><br>
+
   </div><br><br><br>
  
 +
<br>
  
 +
<h2><b>P R O J E C T</b>  SAFETY</h2>
  
−
<h2>PROJECT SAFETY</h2> 
+
<br>  
 
            
 
            
−
<p>We are growing two different strains of S. elongatus: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to A. platensis, we plan to use PCC 7002, which requires external B12 for survival, as a growth reference for the detection of vitamin B12. PCC 7942 was obtained from the University of California, San Diego (UCSD) iGEM team and PCC 7002 was obtained from Dr. Donald Bryant at Pennsylvania State University.<p>
+
<!--<p>We are growing two different strains of <i>S. elongatus</i>: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to <i>A. platensis</i>, we plan to use PCC 7002, which requires external B<sub>12</sub> for survival, as a growth reference for the detection of vitamin B<sub>12</sub>. PCC 7942 was obtained from the University of California, San Diego (UCSD) iGEM team and PCC 7002 was obtained from Dr. Donald Bryant at Pennsylvania State University.</p>-->
  
−
<h3>Vitamin B-12 Safety</h3>
+
<h3>VITAMIN B<sub>12</sub></h3>
 +
 
 +
<br>
 +
 
 +
<h4 style="text-align: left;"><b>G E N E S</b>  U S E D:</h4>
 +
 
 +
<br>
 +
 
 +
<p style="padding-left: 30px; padding-right: 30px;"><b>I. <i>ssuE</i></b> --> produces FMN reductase<br><br>
 
   
 
   
−
<p>G E N E S  U S E D:<br>
+
          Organism: <i>Synechoccocous elongatus</i> strain PCC 7002
−
I. <i>ssuE</i> => produces FMN reductase<br><br>
+
          <br>
 +
          Biosafety <b>Level I</b>
 +
          <br>
 +
          <br>
 
   
 
   
−
           Organism: synechoccocous 7002<br>
+
           The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH<sub>2</sub>). FMNH<sub>2</sub> is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the <i>ssuE</i> gene<sup>[19,20]</sup>.
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           Biosafety Level I<br><br>
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          FMNH<sub>2</sub> + NADP<sup>+</sup> = FMN + NADPH
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The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH2). FMNH2 is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the ssuE gene [citation]. <br><br>
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          <b>II. <i>bluB</i></b> --> 5,6-DMB synthase
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          <br>
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FMNH2 + NADP+ = FMN + NADPH<br><br>
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          <br>
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          Organism: <i>Sinorhizobium meliloti</i> 1021
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          <br>
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          Biosafety <b>Level I</b>
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<h3>ACETAMINOPHEN</h3>
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<h4 style="text-align: left;"><b>G E N E S</b> U S E D:</h4>
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<p style="padding-left: 30px; padding-right: 30px;"><b>I. <i>4ABH</i></b> --> 4-aminophenol<br><br>
 
   
 
   
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II.  <i>bluB</i> => 5,6-DMB synthase<br>
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          Organism: <i>Agaricus bisporus</i>
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           Organism: Sinorhizobium meliloti 1021<br>
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           <br>
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           Biosafety Level I<br>
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           Biosafety <b>Level I</b>
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          <br>
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          <b>II. <i>nhoA</i></b> --> converts 4-aminophenol into acetaminophen
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<h3>Acetaminophen Safety</h3><br><br>
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          <br>
  
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<p>G E N E S  U S E D:<br>
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          Organism: <i>Escherichia coli</i>
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I. <i>4ABH</i> => 4-aminophenol<br><br>
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           <br>
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           Biosafety <b>Level I</b>
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           Organism: Agaricus bisporus<br>
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          <br>
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           Biosafety Level I<br><br>
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           <br>
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           <br>
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II. <i> nhoA</i> => converts 4-aminophenol into acetaminophen<br>
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</p>
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           Organism: Escherichia Coli<br>
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           Biosafety Level I<br><br><br></p>
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<h3><b>T O X I C I T Y</b> TESTS</h3>
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<p>In order to judge whether our project would possibly be safe for consumption, we performed toxicity tests for the acetaminophen pathway in PCC 7942. Varying amounts of acetaminophen were added to liquid culture and the OD of each sample was observed. Liquid culture samples containing 1.32 mM, 6.62 mM, and 13.23 mM of acetaminophen were left to oxidize overnight and the color change was compared with liquid cultures samples containing similar concentrations of 4-aminophenol.</p>
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<p>In order to judge whether our project would possibly be safe for consumption, we performed toxicity tests for the acetaminophen pathway in PCC 7942. Varying amounts of acetaminophen were added to liquid culture and the OD of each sample was observed. Liquid culture samples containing 1.32 mM, 6.62 mM, and 13.23 mM of acetaminophen were left to oxidize overnight and the color change was compared with liquid cultures samples containing similar concentrations of 4-aminophenol.</p> -->
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          <li>[19] R. Caspi, T. Altman, J. M. Dale, K. Dreher, C. A. Fulcher, F. Gilham, P. Kaipa, A. S. Karthikeyan, A. Kothari, M. Krummenacker, M. Latendresse, L. A. Mueller, S. Paley, L. Popescu, A. Pujar, A. G. Shearer, P. Zhang, and P. D. Karp, “The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of pathway/genome databases,” Nucleic Acids Research, vol. 38, pp. D473–D479, Jan. 2010.</li>
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          <li>[20] “ssuE - FMN reductase (NADPH) - Escherichia coli (strain K12) - ssuE gene & protein.”</li>
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Latest revision as of 23:58, 1 November 2017


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SAFETY



When choosing an organism to help us produce acetaminophen and vitamin B12 we sought out an FDA-approved microbe; however, we resorted to using a non-toxic cyanobacteria, Synechococcus elongatus strain PCC 7942, as a model for Arthrospira platensis. A. platensis is a generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.

L A B SAFETY






Proper PPE for BSL-1 organisms are worn in our laboratory. Within the lab space, every person is expected to be wearing gloves, long pants, close-toed shoes, a lab coat, and protective goggles. All personnel also must be accompanied with one other person so that there is no one alone in lab.





P R O J E C T SAFETY


VITAMIN B12


G E N E S U S E D:


I. ssuE --> produces FMN reductase

Organism: Synechoccocous elongatus strain PCC 7002
Biosafety Level I

The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH2). FMNH2 is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the ssuE gene[19,20].

FMNH2 + NADP+ = FMN + NADPH

II. bluB --> 5,6-DMB synthase

Organism: Sinorhizobium meliloti 1021
Biosafety Level I



ACETAMINOPHEN


G E N E S U S E D:


I. 4ABH --> 4-aminophenol

Organism: Agaricus bisporus
Biosafety Level I

II. nhoA --> converts 4-aminophenol into acetaminophen

Organism: Escherichia coli
Biosafety Level I



  • [19] R. Caspi, T. Altman, J. M. Dale, K. Dreher, C. A. Fulcher, F. Gilham, P. Kaipa, A. S. Karthikeyan, A. Kothari, M. Krummenacker, M. Latendresse, L. A. Mueller, S. Paley, L. Popescu, A. Pujar, A. G. Shearer, P. Zhang, and P. D. Karp, “The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of pathway/genome databases,” Nucleic Acids Research, vol. 38, pp. D473–D479, Jan. 2010.
  • [20] “ssuE - FMN reductase (NADPH) - Escherichia coli (strain K12) - ssuE gene & protein.”


    • S P O N S O R S
    S O C I A L
    C O N T A C T
    Jack Baskin School of Engineering
    University of California, Santa Cruz
    1156 High Street
    Santa Cruz, California, 95064