Difference between revisions of "Team:CGU Taiwan/Safety"

 
(4 intermediate revisions by one other user not shown)
Line 2: Line 2:
 
{{CGU_Taiwan/cgustyle}}
 
{{CGU_Taiwan/cgustyle}}
 
{{CGU_Taiwan/notebookstyle}}
 
{{CGU_Taiwan/notebookstyle}}
−
 
 
<html>
 
<html>
 
<head>
 
<head>
 
<meta charset="UTF-8">
 
<meta charset="UTF-8">
−
<title>iGem CGU_Taiwan 2017 - Notebook</title>
+
<title>iGem CGU_Taiwan 2017 - DEMONSTRATION</title>
 
<link href="cgustyle.css" rel="stylesheet" type="text/css" />
 
<link href="cgustyle.css" rel="stylesheet" type="text/css" />
−
<link href="notebookstyle.css" rel="stylesheet" type="text/css" />
 
 
<script type="text/javascript" src="jquery-1.7.2.min.js"></script>
 
<script type="text/javascript" src="jquery-1.7.2.min.js"></script>
 
</head>
 
</head>
  
−
<body bgcolor="white">
+
<body bgcolor="White">
  
 
<!-- nav start -->
 
<!-- nav start -->
 
<div class="head"></div>
 
<div class="head"></div>
−
 
 
<nav>
 
<nav>
 
<ul>
 
<ul>
Line 27: Line 24:
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Results">RESULTS</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Results">RESULTS</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/InterLab">INTERLAB</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/InterLab">INTERLAB</a>
−
       <a href="#">PROOF</a>
+
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Demonstrate">PROOF AND DEMONSTRATION</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Model">MODEL</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Model">MODEL</a>
−
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Parts">PART</a>    
+
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Parts">PARTS</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/Future">FUTURE WORK</a>     
 
       </div>
 
       </div>
 
   </li>
 
   </li>
Line 36: Line 34:
 
       <div class="dropdown-content">
 
       <div class="dropdown-content">
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Notebook">LAB NOTE</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Notebook">LAB NOTE</a>
−
       <a href="#">PROTOCOLS</a>
+
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Protocols">PROTOCOLS</a>
−
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Safety">SATEFY</a>
+
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Safety">SAFETY</a>
 
       </div>
 
       </div>
 
   </li>
 
   </li>
 
   |
 
   |
−
   <li class="dropdown"><a href="Team.html">TEAM</a>
+
   <li class="dropdown"><a href="https://2017.igem.org/Team:CGU_Taiwan/Team">TEAM</a>
 
       <div class="dropdown-content">
 
       <div class="dropdown-content">
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Attributions">ATTRIBUTION</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Attributions">ATTRIBUTION</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Team">MEMBER</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Team">MEMBER</a>
−
       <a href="#">SPONSOR</a>
+
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Acknowledgement">ACKNOWLEDGEMENT</a>
−
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Collaborations">COLLBORATION</a>
+
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Collaborations">COLLABORATION</a>
 
       </div>
 
       </div>
 
   </li>
 
   </li>
Line 55: Line 53:
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/HP/Gold_Integrated">INTERGRATED AND GOLD</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/HP/Gold_Integrated">INTERGRATED AND GOLD</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Engagement">PUBLIC ENGAGEMENT</a>
 
       <a href="https://2017.igem.org/Team:CGU_Taiwan/Engagement">PUBLIC ENGAGEMENT</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/TWConference">2017 iGEM TAIWAN CONFERENCE</a>
 
       </div>
 
       </div>
 
   </li>
 
   </li>
 
   |
 
   |
−
  <li><a href="">AWARD</a></li>
+
<li class="dropdown2"><a href="">AWARD</a>
−
    
+
      <div class="dropdown2-content">
 +
     
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/Entrepreneurship">ENTREPRENEURSHIP</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/Hardware">HARDWARE</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/Measurement">MEASUREMENT</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/Model">MODEL</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/InterLab">INTERLAB</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/HP/Gold_Integrated">INTERGRATED AND GOLD</a>
 +
      <a href="https://2017.igem.org/Team:CGU_Taiwan/Engagement">PUBLIC ENGAGEMENT</a>
 +
      </div>
 +
  </li>
 +
   <li> &nbsp;&nbsp;&nbsp; </li>
 
</ul>
 
</ul>
 
</nav>
 
</nav>
 +
  
 
<script type="text/javascript">
 
<script type="text/javascript">
Line 74: Line 85:
 
         $('.head').css('transform', 'translateX(500px) ');
 
         $('.head').css('transform', 'translateX(500px) ');
 
     }
 
     }
−
 
+
−
if ( $(window).scrollTop() >= 0 ) {
+
if ( $(window).scrollTop() >= 0 ) {
 
         var d = "deg)";
 
         var d = "deg)";
 
         var rot = "rotate(";
 
         var rot = "rotate(";
−
         var rD = Math.floor(($(window).scrollTop())/200) * 90;
+
         var rD = ($(window).scrollTop()) * 0.5;
 
         var rDC = rD.toString();
 
         var rDC = rD.toString();
 
         rot = rot.concat(rDC.concat(d));
 
         rot = rot.concat(rDC.concat(d));
Line 85: Line 96:
 
         $('.square').css('transform', 'none');
 
         $('.square').css('transform', 'none');
 
     }
 
     }
−
 
+
 +
 
})
 
})
 
;
 
;
Line 93: Line 105:
 
<!--background animation start -->
 
<!--background animation start -->
 
<div class="leftMenuBack"></div>
 
<div class="leftMenuBack"></div>
−
<div class="leftMenu">
+
<div class="square"></div>
−
<ul>
+
−
<li class="left1"><a href="#biolist"></a></li>
+
−
<li class="left2"><a href="#modellist"></a></li>
+
−
<li class="left3"><a href="#engineeringlist"></a></li>
+
−
<li class="left4"><a href="#marketinglist"></a></li>
+
−
</ul>
+
−
</div>
+
−
<div class="square"></div>  
+
 
<!--background animation end -->
 
<!--background animation end -->
  
−
<!-- safety start -->
+
<!-- Team start -->
−
<div class="notebook">
+
<div class="description" style="text-align:center">
−
<div class="notebooktitle"><h2>NOTEBOOK | SAFETY</h2></div>
+
<h1 id="pBio"><br>Safety</h1>
−
</div>
+
<h2>1.Brief introduction of the project</h2>
−
<!-- safety end -->
+
  <p style="font-size:120%;">
 +
      Transgenic yeast is designed to be activated by light, being able to secrete out the enzyme which mainly catalyze cellulolysis. Instead of using chemicals, we introduced a new model to detach the ink particle from paper fiber specifically.
 +
</p>
 +
<h2>2.The organisms we used</h2>
 +
  <p style="font-size:120%;">
 +
      east (Saccharomyces cerevisiae), E. coli (DH5α)
 +
</p>
 +
<h2>3.The parts we used</h2>
 +
  <p style="font-size:120%;">
 +
(1) Three enzymes for cellulolysis
 +
endo-beta-1,4-glucanase B, endo-1,4-beta-xylanase A and lipase ORF sequence are from the genome of Aspergillus oryzae, Neocallimastix patriciarum and Rhizopus niveus respectively.
 +
(2) Ste12 promoter and protein of positive up-regulation
 +
We retrieved the sequence of ste12 promoter from upstream 500bp of a pheromone-related pathway transcription factor of Saccharomyces cerevisiae. And we directly acquired the sequence of ste12 protein which is the product of this pathway.
 +
(3) Improved GAL4 based yeast light-switchable promoter system
 +
We adapted the sequence of GAL4 based yeast light-switchable promoter system from BBa_K801042, which is originally designed by Dong-Jiunn Jeffery TRUONG and the group iGEM12_TU_Munich.
 +
</p>
 +
<h2>4.The operation risks during experiments and their solution</h2>
 +
  <p style="font-size:120%;">
 +
(1) Molecular cloning of E. coli
 +
In order to make DNA visible on agarose gels, ethidium bromide was used for staining. It could insert itself between double stranded DNA of samples as well as ours and act as a mutagen which affects DNA biological processes, such as DNA replication and transcription. To prevent skin contact, we not only used protective gloves but also changed it frequently to avoid any ethidium bromide left over. All materials that contact with ethidium bromide were considered polluted and then they should be separately disposed of.
 +
(2) Molecular cloning of yeast
 +
To select the transgenic yeast we interested, the success one will grow up with drug resistance. As we have selected out the yeast wanted, the remaining medium is into the discard, but there is still great amount of yeast in it. If transgenic yeast which has drug resistance flows out, it might become booming and harmful in the nature. To deter from this happening, we added bleach into the medium or sent to sterilization for security.
 +
(3) Test of enzyme activity
 +
We detected the amount of total carbonhydrate with concentrated sulfuric acid and phenol sulfuric acid. They’re both strong reagent which may cause serious dehydration toward our skin. Besides the lab coat and protective gloves, we even wore goggles to prevent from any possible contact.
 +
</p>
 +
<h2>5.Any safety issues raised from the devices?</h2>
 +
  <p style="font-size:120%;">
 +
    Environmental issue
 +
As the device will spray the transgenic yeast onto a paper, there might be a question whether it’s going to stay alive or not after the reprocess. The answer is negative because one of the process will heat to high temperature which kills the yeast for sure. Second, since our device isn’t sealed up completely, the yeast coming out from spray-head will be hard to be removed from the border of box and it might contaminate the gene pool in the environment if they’re not sterilized from the abandon boxes. Thus, we should treat the device a disinfectant to stay clean and safe after use.
 +
</p>
  
−
<!-- javascript code start -->
+
    <br><br>
  
−
<!-- javascript code end -->
+
</div>
 +
<!-- Team end -->
  
 
<!-- footer start -->
 
<!-- footer start -->
−
<footer align="right">   
+
<footer align="center">   
 
<ul>
 
<ul>
−
<li><a href="http://www.cgu.edu.tw"><img src="https://static.igem.org/mediawiki/2017/1/13/Cgucopyright0915.png" width="40%"></a></li>
+
<li><a href="http://www.cgu.edu.tw"><img src="https://static.igem.org/mediawiki/2017/d/d6/Cguwikifooter.png" width="100%"></a></li>
 
</ul>     
 
</ul>     
 
</footer>
 
</footer>

Latest revision as of 03:17, 16 December 2017

iGem CGU_Taiwan 2017 - DEMONSTRATION


Safety

1.Brief introduction of the project

Transgenic yeast is designed to be activated by light, being able to secrete out the enzyme which mainly catalyze cellulolysis. Instead of using chemicals, we introduced a new model to detach the ink particle from paper fiber specifically.

2.The organisms we used

east (Saccharomyces cerevisiae), E. coli (DH5α)

3.The parts we used

(1) Three enzymes for cellulolysis endo-beta-1,4-glucanase B, endo-1,4-beta-xylanase A and lipase ORF sequence are from the genome of Aspergillus oryzae, Neocallimastix patriciarum and Rhizopus niveus respectively. (2) Ste12 promoter and protein of positive up-regulation We retrieved the sequence of ste12 promoter from upstream 500bp of a pheromone-related pathway transcription factor of Saccharomyces cerevisiae. And we directly acquired the sequence of ste12 protein which is the product of this pathway. (3) Improved GAL4 based yeast light-switchable promoter system We adapted the sequence of GAL4 based yeast light-switchable promoter system from BBa_K801042, which is originally designed by Dong-Jiunn Jeffery TRUONG and the group iGEM12_TU_Munich.

4.The operation risks during experiments and their solution

(1) Molecular cloning of E. coli In order to make DNA visible on agarose gels, ethidium bromide was used for staining. It could insert itself between double stranded DNA of samples as well as ours and act as a mutagen which affects DNA biological processes, such as DNA replication and transcription. To prevent skin contact, we not only used protective gloves but also changed it frequently to avoid any ethidium bromide left over. All materials that contact with ethidium bromide were considered polluted and then they should be separately disposed of. (2) Molecular cloning of yeast To select the transgenic yeast we interested, the success one will grow up with drug resistance. As we have selected out the yeast wanted, the remaining medium is into the discard, but there is still great amount of yeast in it. If transgenic yeast which has drug resistance flows out, it might become booming and harmful in the nature. To deter from this happening, we added bleach into the medium or sent to sterilization for security. (3) Test of enzyme activity We detected the amount of total carbonhydrate with concentrated sulfuric acid and phenol sulfuric acid. They’re both strong reagent which may cause serious dehydration toward our skin. Besides the lab coat and protective gloves, we even wore goggles to prevent from any possible contact.

5.Any safety issues raised from the devices?

Environmental issue As the device will spray the transgenic yeast onto a paper, there might be a question whether it’s going to stay alive or not after the reprocess. The answer is negative because one of the process will heat to high temperature which kills the yeast for sure. Second, since our device isn’t sealed up completely, the yeast coming out from spray-head will be hard to be removed from the border of box and it might contaminate the gene pool in the environment if they’re not sterilized from the abandon boxes. Thus, we should treat the device a disinfectant to stay clean and safe after use.