Difference between revisions of "Team:WHU-China/Safety"

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<p> IGEM-WHU </p>
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<nav class="navbar navbar-expand-lg navbar-light bg-light">
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<h1> Safety </h1>
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<div class="collapse navbar-collapse justify-content-center" id="navbarSupportedContent">
<p>Please visit the <a href="https://2018.igem.org/Safety">Safety Hub</a> to find this year's safety requirements & deadlines, and to learn about safe & responsible research in iGEM.</p>
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                                <li><a href="https://2018.igem.org/Team:WHU-China/Safety">Safety</a></li>
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<p>On this page of your wiki, you should write about how you are addressing any safety issues in your project. The wiki is a place where you can <strong>go beyond the questions on the safety forms</strong>, and write about whatever safety topics are most interesting in your project. (You do not need to copy your safety forms onto this wiki page.)</p>
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                      <li><a href="https://2018.igem.org/Team:WHU-China/Public_Engagement">Education engagement</a></li>
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                <li><a href="https://2018.igem.org/Team:WHU-China/Hardware">Hardware</a></li>
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</nav>
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</header>
  
  
<div class="column two_thirds_size">
 
<h3>Safe Project Design</h3>
 
  
<p>Does your project include any safety features? Have you made certain decisions about the design to reduce risks? Write about them here! For example:</p>
 
  
<ul>
 
<li>Choosing a non-pathogenic chassis</li>
 
<li>Choosing parts that will not harm humans / animals / plants</li>
 
<li>Substituting safer materials for dangerous materials in a proof-of-concept experiment</li>
 
<li>Including an "induced lethality" or "kill-switch" device</li>
 
</ul>
 
  
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<div class="c_row">
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<br/><br/><br/><br/><br/><br/>
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<h1>Safety</h1>
 +
<br />
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<p style="font-size:22px;">Security is an important issue for all the teams especially environment track. Actually, we pay great attention to safety issues, from laboratory safety to design project safety. We are cautious at every step to ensure that there are no problems with safety.</p>
 +
<br />
 +
<br />
 +
<br />
 +
<h1>1.Project safety</h1>
 
</div>
 
</div>
  
<div class="column third_size">
 
<h3>Safe Lab Work</h3>
 
  
<p>What safety procedures do you use every day in the lab? Did you perform any unusual experiments, or face any unusual safety issues? Write about them here!</p>
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<div class="c_row" style="text-align:center;">
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<img src="https://static.igem.org/mediawiki/2018/3/3f/T--WHU-China--wiki-safety_main1.png" style="width:70%;">
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</div>
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<div class="c_row">
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<p style="font-size:22px;">In our project, we will put our Noah’s Ark into the environment water, and our engineered bacteria will make contact with water in turbine to accumulate CC. It seems that we have encountered the same problems as other teams - GMO leakage.
 +
However, we have set up multiple lines of defense to prevent bacteria from escaping from our ark.</p>
 +
<br />
 +
<h2>(1). Biofilm</h2>
 +
<br />
 +
<p style="font-size:22px;">Most bacteria are tightly wrapped by biofilms, owing to the EPS secreted by algae. What’s more, the second plasmid can anchor the bacteria  tightly onto biofilm. These together allow them to be fixed on the track and unable to enter the liquid environment inside the turbine. </p>
 +
<br />
 +
<h2>(2). microfiltration membrane</h2>
 +
<br />
 +
<p style="font-size:22px;">Water and the compounds dissolved in it can enter and exit the turbine freely, but cells can’t. This is because we add multiple microfiltration membrane to the water inlet and outlet of the turbine. Each single layer of filter can ensure that the bacteria can not pass. We screened the microfiltration membrane carefully and finally chose the Mixed fiber microfiltration membrane.</p>
 +
<br />
 +
<h3>1).what is microfiltration membrane</h3>
 +
<br />
 +
<p style="font-size:22px;">In the application of membrane separation technology, microfiltration membrane is the most widely used membrane variety. It is widely used in many fields such as scientific research, food testing, chemical industry, nanotechnology, energy and environmental protection. It is mainly used for the filtration of aqueous solutions, so it is also called aqueous film.
 +
</p>
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</div>
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<div class="c_row">
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<div style="float:left;">
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  <table border="2" cellpadding="0" cellspacing="0" width="458" bordercolor="#FFFFFF"; style="font-size:15px;">
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  <col width="229" span="2" />
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  <tr bgcolor="#4472C4">
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    <td width="229"><p style="font-size:16px;color:white;">Aperture</p></td>
 +
    <td width="229"><p style="font-size:16px;color:white;">Application</p></td>
 +
  </tr>
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  <tr>
 +
    <td width="229" bgcolor="#4472C4"><p style="font-size:16px;color:white;">3.0-10.0μm</p></td>
 +
    <td width="229" bgcolor="#CFD5EA"><p style="font-size:16px;">Security filtering before RO desalination</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="229" bgcolor="#4472C4"><p style="font-size:16px;color:white;">0.6-0.8μm</p></td>
 +
    <td width="229" bgcolor="#E9EBF5"><p style="font-size:16px;">High-dose injection, particle filtration in large infusion, clarification filtration of    beer, beverage filtration, oil photoresist, spray solvent, etc.</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="229" bgcolor="#4472C4"><p style="font-size:16px;color:white;">0.45μm</p></td>
 +
    <td width="229" bgcolor="#CFD5EA"><p style="font-size:16px;">High-purity water terminal filtration in the electronics industry, filtration of MOS    reagents</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="229" bgcolor="#4472C4"><p style="font-size:16px;color:white;">0.2μm</p></td>
 +
    <td width="229" bgcolor="#E9EBF5"><p style="font-size:16px;">Sterilization and filtration of liquid medicine, biological agents and heat-sensitive    fluids, ultra-pure water terminal filtration in the electronics industry, and clarification filtration of low-grade wines</p></td>
 +
  </tr>
 +
</table>
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  </div>
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<div style="float:right;">
 +
  <table border="2" bordercolor="#FFFFFF"  cellpadding="0" cellspacing="0" width="423">
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  <col width="211" span="2" />
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  <tr bgcolor="#4472C4">
 +
    <td width="211"><p style="font-size:16px;color:white;">Name</p></td>
 +
    <td width="211"><p style="font-size:16px;color:white;">Usage</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="211" bgcolor="#4472C4"><p style="font-size:16px;color:white;">Cellulose membrane</p></td>
 +
    <td width="211" bgcolor="#CFD5EA"><p style="font-size:16px;">Preparation of nanofiltration  membrane, microfiltration membrane</p>
 +
    <p style="font-size:16px;">Polyamides:</p>
 +
    <p style="font-size:16px;">&nbsp;</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="211" bgcolor="#4472C4"><p style="font-size:16px;color:white;">Polyamides</p></td>
 +
    <td width="211" bgcolor="#E9EBF5"><p style="font-size:16px;">&nbsp;</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="211" bgcolor="#4472C4"><p style="font-size:16px;color:white;">Polysulfones</p></td>
 +
    <td width="211" bgcolor="#CFD5EA"><p style="font-size:16px;">mostly used in ultrafiltration membranes, gas separation membranes</p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="211" bgcolor="#4472C4"><p style="font-size:16px;color:white;">Inorganic membrane</p></td>
 +
    <td width="211" bgcolor="#E9EBF5"><p style="font-size:16px;">used as a catalytic carrier</p>
 +
      <p style="font-size:16px;">&nbsp;</p></td>
 +
  </tr>
 +
</table>
 +
 +
  </div>
 +
</div>
 +
<div class="c_row" style="clear:both">
 +
<br />
 +
<h3>2). mixed fiber microfiltration membrane</h3>
 +
<br />
 +
<p style="font-size:22px;">Since E.coli has a diameter of 0.5-1.3 μm, finally we chose mixed fiber microfiltration membrane(0.22 μm) , Generally referred to nitric acid-acetic acid mixed cellulose ester microporous membrane, is widely used in many fields such as Pharmaceutical industry, Electronics industry, Public health, Food industry and Sterility testing.</p>
 +
<br />
 +
<p style="font-size:22px;">Multiple layers of filtration membrane gives further insurance to prevent the leakage.</p>
 +
<br />
 +
<h2>(3).” UV tunnel”</h2><br />
 +
<p style="font-size:22px;">Just like the laser tunnel in the movie: Resident Evil, in our design, an ultraviolet device is added at the exit of the turbine end to prevent leakage. Once the bacteria escapes to the exit, they will be killed by ultraviolet rays.</p>
 +
</div>
  
 +
<div class="c_row" style="text-align:center;">
 +
<img src="https://static.igem.org/mediawiki/2018/c/cc/T--WHU-China--wiki-safety_main2.png" style="width:50%;">
 +
</div>
  
<h3>Safe Shipment</h3>
+
<div class="c_row">
 +
<br />
 +
<p style="font-size:22px;">Thus, if the bacteria want to escape, it must go through a long way to the outside. And if that happens, congratulations! Because bacteria will then commit a suicide—that is our last defense!</p>
 +
<br />
 +
<h2>(4) Kill switch</h2>
 +
<br />
 +
<p style="font-size:22px;">To prevent bacterial leakage, we designed two toxin-antitoxin pathways to ensure that bacteria will commit a suicide once they leave the environment we provide.  In our experiment, one of our pathway worked—”ParD/E”, to see more experiment details you can go to “laboratory-molecular biology lab-experiment” to see how bacteria will be killed.</p>
 +
<br />
 +
</div>
  
<p>Did you face any safety problems in sending your DNA parts to the Registry? How did you solve those problems?</p>
+
<div class="c_row" style="text-align:center;">
 +
<img src="https://static.igem.org/mediawiki/2018/8/8a/T--WHU-China--wiki-safety_main3.png" style="width:50%;">
 
</div>
 
</div>
  
 +
<div class="c_row">
 +
<br />
 +
<p style="font-size:22px;">What’s more, there is another design to provide a more reliable method—a cross-overdox antitoxin plasmid system ensures that the loss of any one of these plasmids can cause death, which can solve the risk of possible horizontal gene transfer.</p>
 +
<br />
 +
</div>
  
 +
<div class="c_row" style="text-align:center;">
 +
<img src="https://static.igem.org/mediawiki/2018/b/bd/T--WHU-China--wiki-safety_main4.png" style="width:60%;">
 +
</div>
 +
 +
<div class="c_row">
 +
<br />
 +
<p style="font-size:22px;">Since we have two plasmids, we want to design like this to prevent the HGT happening because loss or gain of any one plasmid can lead to suicide by the toxic on it.</p>
 +
<br />
 +
<p style="font-size:22px;">Conclusion: Because of time limitation, we had not verified the experiments on UV and microfiltration membranes. The drop rate of bacteria on the biofilm and kill switch had been partially verified. We spent a lot of energy on design considerations and hardware safety considerations. If it could be put into application, we can guarantee that the above defenses can prevent leakage problems as much as possible, and more security experiments will continue to be done.</p>
 +
<br/><br/>
 +
<h1>2.Lab safety</h1>
 +
<p style="font-size:22px;">We learnt laboratory safety before entering the lab. Our PI trained us in aspect of basic chemical safety and biosafety.
 +
We followed all precautions, including wearing gloves at all times, tightly controlling lab accessibility, avoiding all food and drink in the lab, enforcing hand washing with 70% alcohol before leaving the lab to ensure the utmost safety and so on.
 +
The pictures below shows the overview of our lab (scientific training lab) in Wuhan University. We have divisions of experimental areas and all chemical reagents are classified and well reserved.</p>
 +
<br/><br/>
 +
 +
</div>
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 +
<div class="c_row">
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<div style="float:left;width:300px;">
 +
  <img src="https://static.igem.org/mediawiki/2018/e/ee/T--WHU-China--wiki-safety_main5.jpg">
 +
  </div>
 +
<div style="float:left;width:300px;margin-left:50px;">
 +
  <img src="https://static.igem.org/mediawiki/2018/b/be/T--WHU-China--wiki-safety_main6.jpg">
 +
  </div>
 +
  <div style="float:left;width:300px;margin-left:50px;">
 +
  <img src="https://static.igem.org/mediawiki/2018/c/c4/T--WHU-China--wiki-safety_main7.jpg">
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  </div>
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Latest revision as of 03:26, 18 October 2018

Safety







Safety


Security is an important issue for all the teams especially environment track. Actually, we pay great attention to safety issues, from laboratory safety to design project safety. We are cautious at every step to ensure that there are no problems with safety.




1.Project safety

In our project, we will put our Noah’s Ark into the environment water, and our engineered bacteria will make contact with water in turbine to accumulate CC. It seems that we have encountered the same problems as other teams - GMO leakage. However, we have set up multiple lines of defense to prevent bacteria from escaping from our ark.


(1). Biofilm


Most bacteria are tightly wrapped by biofilms, owing to the EPS secreted by algae. What’s more, the second plasmid can anchor the bacteria tightly onto biofilm. These together allow them to be fixed on the track and unable to enter the liquid environment inside the turbine.


(2). microfiltration membrane


Water and the compounds dissolved in it can enter and exit the turbine freely, but cells can’t. This is because we add multiple microfiltration membrane to the water inlet and outlet of the turbine. Each single layer of filter can ensure that the bacteria can not pass. We screened the microfiltration membrane carefully and finally chose the Mixed fiber microfiltration membrane.


1).what is microfiltration membrane


In the application of membrane separation technology, microfiltration membrane is the most widely used membrane variety. It is widely used in many fields such as scientific research, food testing, chemical industry, nanotechnology, energy and environmental protection. It is mainly used for the filtration of aqueous solutions, so it is also called aqueous film.

Aperture

Application

3.0-10.0μm

Security filtering before RO desalination

0.6-0.8μm

High-dose injection, particle filtration in large infusion, clarification filtration of beer, beverage filtration, oil photoresist, spray solvent, etc.

0.45μm

High-purity water terminal filtration in the electronics industry, filtration of MOS reagents

0.2μm

Sterilization and filtration of liquid medicine, biological agents and heat-sensitive fluids, ultra-pure water terminal filtration in the electronics industry, and clarification filtration of low-grade wines

Name

Usage

Cellulose membrane

Preparation of nanofiltration membrane, microfiltration membrane

Polyamides:

 

Polyamides

 

Polysulfones

mostly used in ultrafiltration membranes, gas separation membranes

Inorganic membrane

used as a catalytic carrier

 


2). mixed fiber microfiltration membrane


Since E.coli has a diameter of 0.5-1.3 μm, finally we chose mixed fiber microfiltration membrane(0.22 μm) , Generally referred to nitric acid-acetic acid mixed cellulose ester microporous membrane, is widely used in many fields such as Pharmaceutical industry, Electronics industry, Public health, Food industry and Sterility testing.


Multiple layers of filtration membrane gives further insurance to prevent the leakage.


(3).” UV tunnel”


Just like the laser tunnel in the movie: Resident Evil, in our design, an ultraviolet device is added at the exit of the turbine end to prevent leakage. Once the bacteria escapes to the exit, they will be killed by ultraviolet rays.


Thus, if the bacteria want to escape, it must go through a long way to the outside. And if that happens, congratulations! Because bacteria will then commit a suicide—that is our last defense!


(4) Kill switch


To prevent bacterial leakage, we designed two toxin-antitoxin pathways to ensure that bacteria will commit a suicide once they leave the environment we provide. In our experiment, one of our pathway worked—”ParD/E”, to see more experiment details you can go to “laboratory-molecular biology lab-experiment” to see how bacteria will be killed.



What’s more, there is another design to provide a more reliable method—a cross-overdox antitoxin plasmid system ensures that the loss of any one of these plasmids can cause death, which can solve the risk of possible horizontal gene transfer.



Since we have two plasmids, we want to design like this to prevent the HGT happening because loss or gain of any one plasmid can lead to suicide by the toxic on it.


Conclusion: Because of time limitation, we had not verified the experiments on UV and microfiltration membranes. The drop rate of bacteria on the biofilm and kill switch had been partially verified. We spent a lot of energy on design considerations and hardware safety considerations. If it could be put into application, we can guarantee that the above defenses can prevent leakage problems as much as possible, and more security experiments will continue to be done.



2.Lab safety

We learnt laboratory safety before entering the lab. Our PI trained us in aspect of basic chemical safety and biosafety. We followed all precautions, including wearing gloves at all times, tightly controlling lab accessibility, avoiding all food and drink in the lab, enforcing hand washing with 70% alcohol before leaving the lab to ensure the utmost safety and so on. The pictures below shows the overview of our lab (scientific training lab) in Wuhan University. We have divisions of experimental areas and all chemical reagents are classified and well reserved.