Difference between revisions of "Team:UNIK Copenhagen/Green Lab"

 
(30 intermediate revisions by 4 users not shown)
Line 2: Line 2:
 
<html>
 
<html>
 
<style>
 
<style>
 +
  
 
#imagebox{
 
#imagebox{
−
font-size:10px;
+
      font-size:11px;
 +
      float:left;
 +
      width:900px;
 
}
 
}
 +
 
.imageboxshadow img{
 
.imageboxshadow img{
 
box-shadow: 0px 0px 5px #888888;
 
box-shadow: 0px 0px 5px #888888;
 
-webkit-box-shadow: 0px 0px 5px #888888;
 
-webkit-box-shadow: 0px 0px 5px #888888;
 
-moz-box-shadow: 0px 0px 5px #888888;
 
-moz-box-shadow: 0px 0px 5px #888888;
 +
}
 +
 +
.container img{
 +
box-shadow: 0px 0px 5px #888888;
 +
-webkit-box-shadow: 0px 0px 5px #888888;
 +
-moz-box-shadow: 0px 0px 5px #888888;
 +
}
 +
 +
div.container {
 +
      display:inline-block;
 +
      clear: left;
 +
      width:380px;
 +
    }
 +
 +
p{
 +
text-align:justify;
 +
}
 +
 +
.container p.{
 +
text-align:center;
 +
font-size: 11px;
 
}
 
}
  
Line 17: Line 42:
 
<h1><font color="green">Green</font> Lab</h1>
 
<h1><font color="green">Green</font> Lab</h1>
  
−
We will make a stable and transient transformation of moss, Psychometrilla patens. We will make two linear gene constructs for stable transformation in moss. One construct is going to contain a gene encoding an antifreeze protein and the other construct will contain a stilbene synthase-gene (STS), making the final enzyme in the resveratrol pathway. Both constructs contain regions homologous to the moss genome, so they will integrate via homologous recombination in moss. The constructs furthermore contains the ZmUbi-promoter, resistance gene and YFP, to confirm transformation. We will also make a transient moss transformation with a vector containing the ZmUbi-promoter followed by YFP, to confirm the function of the promoter in moss.  
+
<br>
 +
 
 +
<div id="imagebox">
 +
<div class="imageboxshadow">
 +
<img src="https://static.igem.org/mediawiki/2015/6/67/UNIK_copenhagen_greenlabteam.jpg" width=80% style="margin: 0px 0px 0px 100px"></div>
 +
<p style="font-size:10.5px; margin: 4px 170px 0px 100px">Green Lab team</p> <br><br></div>
 +
 
 
<br><br>
 
<br><br>
  
 +
<H3>What is Green Lab About?</h3>
 +
<p>
 +
Simply put, the aim of Green Lab is to create transgenic moss. Or to be specific, we want to transform the bryophyte species, <i>Physcomitrella patens</i>. But why would you want to use moss for laboratory experiments instead of the usual chassis organism, like bacteria or yeast? The short answer is: Moss can do awesome things. The longer answer is the following: Moss is a photosynthetic organism and thus produce oxygen, where yeast and (most) bacteria are heterotrophic organism that cannot produce oxygen. Being able to do photosynthesis and produce oxygen, could be very useful on a space mission.
 +
 +
<br><br>
 +
 +
Moss is called "the yeast of plant biology" since it - like yeast - is able to do homologous recombination [1]⁠. Moss has already been used extensively as a model organism in plant biology and has been used as a bioreactor for many different compounds [2]⁠.
 +
Also, moss is a hardy plant. Moss can grow under many conditions, such as in the shade, and high light on the surface of rocks. And for the reasons discussed below, we do need a tough organism for our plans to succeed.</p>
 +
 +
<br>
  
 
<div id="imagebox">
 
<div id="imagebox">
 
<div class="imageboxshadow">
 
<div class="imageboxshadow">
−
<img src="https://static.igem.org/mediawiki/2015/3/3e/UNIK_copenhagen_mossbasement.jpg" width=62%></div>
+
<img src="https://static.igem.org/mediawiki/2015/8/8d/UNIK_Copenhagen_Moss_landscape.png" width=65% style="margin: 0px 0px 0px 142px">
−
Moss growing facility</div>
+
</div>
 +
<p style="font-size:10.5px; margin: 4px 170px 0px 142px">How Mars should look</p>
  
 +
<br>
  
 +
<p>We envision that the astronauts eventually will be able to plant the moss directly on the surface of Mars. If we could make moss survive on Mars, we would in a sense create a useful "harddrive" on which the "software" can be installed. This "software" could be different biopharmaceutical ingredient productions. Thus, we wanted to make <i>P. patens</i> more adapted to the martian environment. There are many factors on Mars that are lethal to living organisms, but as a start we would focus on the temperatures on Mars. The temperatures on Mars can drop well below zero at night and therefore we found a novel antifreeze protein for expression in the moss.
 +
This could be an important first step in designing organisms suited for the martian environment, which can help create a sustainable settlement on Mars.
 +
 +
<br><br>
 +
 +
In principle, the Mars adapted moss could be transformed to produce many of the biopharmaceutical ingredients that astronauts may need on a space mission. As a proof of this concept we wanted our moss to produce Resveratrol.
 +
Resveratrol is a phenolic compound normally found in red wine and is believed to have positive health benefits in mammals [3],[4]. If this compound can be produced, it opens the door for a wide range of other applications, that is essential for space exploration.
 +
This would prove, that different kinds of "software" can be installed on the Mars adapted moss "hardware".</p>
 +
 +
<br>
 +
 +
<div id="imagebox">
 +
<img src="https://static.igem.org/mediawiki/2015/e/e9/UNIK_Copenhagen_Easyidea.jpg" width=550px style="margin: 0px 0px 0px 170px">
 +
<br><br><br>
 +
</div>
 +
 +
<br><br>
 +
 +
<p>However, using moss for experiments comes at a price. If you ever find yourself divided between two choices: watching moss grow or watching paint dry, opt for the latter. Moss grows slower than bacteria and yeast, but faster than most higher plants. Moss is relatively simply to stably transform. After transformation of moss protoplasts, it takes about eight weeks for the moss to form small clumps when grown on a petri dish, thus experiments with moss need to be well planned. </p>
 +
 +
<br><br>
 +
 +
<div class="container">
 +
<img src="https://static.igem.org/mediawiki/2015/3/3e/UNIK_copenhagen_mossbasement.jpg" width=430px>
 +
<p style="font-size:11px">Moss growing facility</p>
 +
</div>
 +
 +
<div class="container">
 +
<img src="https://static.igem.org/mediawiki/2015/3/33/UNIK_Copenhagen_Jonandadam.jpg" width=430px style="margin:0px 0px 0px 80px">
 +
<p style="font-size:11px; margin:4px 0px 0px 80px">Jonathan and Adam working on PCR reactions at the lab</p>
 +
</div>
 +
 +
 +
<br><br><br>
 +
 +
</div>
 +
 +
<p><b>References:</b>
 +
<br>
 +
[1] Kamisugi, Y. et al. The mechanism of gene targeting in Physcomitrella patens: Homologous recombination, concatenation and multiple integration. Nucleic Acids Res. 34, 6205–6214 (2006).<br>
 +
[2] Decker, E. L., Parsons, J. & Reski, R. Glyco-engineering for biopharmaceutical production in moss bioreactors. Front. Plant Sci. 5, 346 (2014).<br>
 +
[3] Bhat KPL, Kosmeder, J. W. & Pezzuto, J. M. Biological effects of resveratrol. Antioxid. Redox Signal. 3, 1041–1064 (2001).<br>
 +
[4] Yang, T., Wang, L., Zhu, M., Zhang, L. & Yan, L. Properties and molecular mechanisms of resveratrol : a review. Div. Chinese Med. 70, 501–506 (2015).
 +
</p>
 +
 +
<br><br>
  
−
<br/>
+
</html>
−
Due to time constraints, we will also transform tobacco and <i>E.coli</i>. We will transform tobacco with the STS-gene, using USER-cloning and Agrobacterium tumefaciens transformation, in order to quickly confirm the functionality of the gene (moss takes a long time to grow!) by measuring the presence of resveratrol with Liquid chromatography-mass spectrometry.
+
−
Furthermore we will transform E. coli with the antifreeze gene which originate from the spruce budworm, to quickly see whether the gene has an effect on temperature sensitivity. We will perform experiments at low temperatures to measure the growth of the bacteria when exposed to cold. We will also make a his-tag version of the antrifreezeprotein, to confirm the presence of the protein by western blotting.
+

Latest revision as of 20:01, 17 September 2015


Green Lab


Green Lab team





What is Green Lab About?

Simply put, the aim of Green Lab is to create transgenic moss. Or to be specific, we want to transform the bryophyte species, Physcomitrella patens. But why would you want to use moss for laboratory experiments instead of the usual chassis organism, like bacteria or yeast? The short answer is: Moss can do awesome things. The longer answer is the following: Moss is a photosynthetic organism and thus produce oxygen, where yeast and (most) bacteria are heterotrophic organism that cannot produce oxygen. Being able to do photosynthesis and produce oxygen, could be very useful on a space mission.

Moss is called "the yeast of plant biology" since it - like yeast - is able to do homologous recombination [1]⁠. Moss has already been used extensively as a model organism in plant biology and has been used as a bioreactor for many different compounds [2]⁠. Also, moss is a hardy plant. Moss can grow under many conditions, such as in the shade, and high light on the surface of rocks. And for the reasons discussed below, we do need a tough organism for our plans to succeed.


How Mars should look


We envision that the astronauts eventually will be able to plant the moss directly on the surface of Mars. If we could make moss survive on Mars, we would in a sense create a useful "harddrive" on which the "software" can be installed. This "software" could be different biopharmaceutical ingredient productions. Thus, we wanted to make P. patens more adapted to the martian environment. There are many factors on Mars that are lethal to living organisms, but as a start we would focus on the temperatures on Mars. The temperatures on Mars can drop well below zero at night and therefore we found a novel antifreeze protein for expression in the moss. This could be an important first step in designing organisms suited for the martian environment, which can help create a sustainable settlement on Mars.

In principle, the Mars adapted moss could be transformed to produce many of the biopharmaceutical ingredients that astronauts may need on a space mission. As a proof of this concept we wanted our moss to produce Resveratrol. Resveratrol is a phenolic compound normally found in red wine and is believed to have positive health benefits in mammals [3],[4]. If this compound can be produced, it opens the door for a wide range of other applications, that is essential for space exploration. This would prove, that different kinds of "software" can be installed on the Mars adapted moss "hardware".







However, using moss for experiments comes at a price. If you ever find yourself divided between two choices: watching moss grow or watching paint dry, opt for the latter. Moss grows slower than bacteria and yeast, but faster than most higher plants. Moss is relatively simply to stably transform. After transformation of moss protoplasts, it takes about eight weeks for the moss to form small clumps when grown on a petri dish, thus experiments with moss need to be well planned.



Moss growing facility

Jonathan and Adam working on PCR reactions at the lab




References:
[1] Kamisugi, Y. et al. The mechanism of gene targeting in Physcomitrella patens: Homologous recombination, concatenation and multiple integration. Nucleic Acids Res. 34, 6205–6214 (2006).
[2] Decker, E. L., Parsons, J. & Reski, R. Glyco-engineering for biopharmaceutical production in moss bioreactors. Front. Plant Sci. 5, 346 (2014).
[3] Bhat KPL, Kosmeder, J. W. & Pezzuto, J. M. Biological effects of resveratrol. Antioxid. Redox Signal. 3, 1041–1064 (2001).
[4] Yang, T., Wang, L., Zhu, M., Zhang, L. & Yan, L. Properties and molecular mechanisms of resveratrol : a review. Div. Chinese Med. 70, 501–506 (2015).