Difference between revisions of "Team:Lethbridge HS/Improve"

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</p>
 
</p>
  
 +
<div class="container-fluid" style="margin-left:10%; margin-right:10%;">
 +
<h2><b>&nbsp;Anthocyanin Constructs</b></h2>
 +
<br>
 +
<h3>Anthocyanin composite part 1</h3>
 +
<p class="center">
 +
<a href="">BBa_K1281101</a>
 +
<br>
 +
<b>T7 Promoter</b> Part <a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>dfr</i></b> Part <a href="http://parts.igem.org/Part:BBa_K1497010">BBa_K1497010</a>
 +
<br>
 +
<b><i>f3h</i></b> Part <a href="http://parts.igem.org/Part:BBa_K1497009">BBa_K1497009</a>
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
</p>
 +
<br>
  
 +
<h3>Anthocyanin composite part 2</h3>
 +
<p class="center">
 +
<a href="">BBa_K1281102</a>
 +
<br>
 +
<b>T7 Promoter</b> Part <a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>ans</i></b> Part <a href="http://parts.igem.org/Part:BBa_K1497002">BBa_K1497002</a>
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
</p>
 +
<br>
 +
 +
<h3>Anthocyanin composite part 3</h3>
 +
<p class="center">
 +
<a href="">BBa_K1281103</a>
 +
<br>
 +
<b>T7 Promoter</b> Part <a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>3gt</i></b> Part <a href="">BBa_K1281003</a>
 +
<br>
 +
<b><i>yad</i>H</b> Part <a href="">BBa_K1281002</a>
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
<br>
 +
<br>
 +
These composite parts come together to complete the pathway from Eriodictyol to Anthocyanin. These constructs are separated due to the size of the genes. It would be too much of a sstrain on the cell to have all of th genes in one or even two plasmids and this is why we needed three separate constructs. These constructs are each submitted
 +
<br>
 +
 +
 +
<div style="text-align:center;"><center><img height=50px class="img-responsive"src="https://static.igem.org/mediawiki/2017/1/15/T--Lethbridge_HS--AnthocyaninPathway.png"></center></div>
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/4/49/T--Lethbridge_HS--Anthocyanincomp1.png"></center></div>
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/9/9f/T--Lethbridge_HS--Anthocyanincomp2.png"></center>
 +
</div>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/0/0c/T--Lethbridge_HS--Anthocyanincomp3.png"></center>
 +
</div>
 +
</p>
 +
<br> 
 +
<br>
 +
 +
<h2><b>&nbsp;Zeaxanthin Construct</b></h2>
 +
<h3>Zeaxanthin composite part</h3>
 +
<p class="center">
 +
<a href="">BBa_K1281301</a>
 +
<br>
 +
<b>T7 Promoter</b> Part <a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>crt</i>Y</b> Part <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_I742154">BBa_I742154</a>.
 +
<br>
 +
<b><i>crt</i>Z</b> Part <a href"http://parts.igem.org/Part:BBa_I742157">BBa_I742157</a>.
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
<br>
 +
<br>
 +
This construct is in the plasmid psB1C3, and will convert our initial molecule Lycopene into our final product, the pigment Zeaxanthin.
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/8/8f/T--Lethbridge_HS--ZeaxanthinPathway.png"></center>
 +
</div>
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/c/c5/T--Lethbridge_HS--ZeaxanthinConstruct.png"></center>
 +
</div>
 +
</p>
 +
<br>
 +
 +
<h2><b>&nbsp;Indigoidine Constructs</b></h2>
 +
<h3>Indigoidine composite part 1</h3>
 +
<p class="center">
 +
<a href="">BBa_K1281201</a>
 +
<br>
 +
<b>T7 Promoter</b> Part<a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>ind</i>B</b> Part <a href="">BBa_K1281001</a>
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
</p>
 +
<br>
 +
 +
<h3>Indigoidine composite part 2</h3>
 +
<p class="center">
 +
<a href="">BBa_K1281202</a>
 +
<br>
 +
<b>T7 Promoter</b> Part<a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>ind</i>C</b> Part <a href="http://parts.igem.org/Part:BBa_K1152013">BBa_K1152013</a>
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
<br>
 +
<br>
 +
These two composite parts come together to convert our initial molecule Glutamine into Indigoidine. The <i>ind</i>B has been shown to increase the yields of Indigoidine as well as it is our original basic part submissions. This construct is part <a href="">BBa_K1281201</a> and part <a href="">BBa_K1281202</a>. We had to split our genes into tow separate composite part submissions as the size of each was too large to allow for them to be in one plasmid.
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/0/02/T--Lethbridge_HS--IndigoidinePathway.png"></center>
 +
</div>
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/2/25/T--Lethbridge_HS--IndigoidineConstruct.png"></center>
 +
</div>
 +
</p>
 +
<br>
 +
<br>
 +
 +
 +
<h2><b>&nbsp;Melanin Construct</b></h2>
 +
<h3>Melanin composite part</h3>
 +
<p class="center">The parts used in our Melanin construct are as follows:
 +
<br>
 +
<b>T7 Promoter</b> Part<a href="http://parts.igem.org/Part:BBa_I712074">BBa_I712074</a>.
 +
<br>
 +
<b>RBS</b> Part <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>.
 +
<br>
 +
<b><i>Mel</i>A</b> Part <a href="http://parts.igem.org/Part:BBa_K274001">BBA_K274001</a>.
 +
<br>
 +
<b>Terminator</b> Part <a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>.
 +
<br>
 +
<br>
 +
This construct allows us to make Melanin out of L-Tyrosine.
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/a/ad/T--Lethbridge_HS--MelaninPathway.png"></center>
 +
</div>
 +
<br>
 +
<div style="text-align:center;"><center><img height="150px"class="img-responsive"src="https://static.igem.org/mediawiki/2017/f/f2/T--Lethbridge_HS--MelaninConstruct.png"></center>
 +
</div>
 +
</p>
 +
<br>
  
  

Revision as of 22:17, 26 October 2017





 Parts Improved

We improved all of the basic parts we used by putting them into the composite parts that we used. The T7 promoter that we had at the beginning of each construct and our use of the T7 system in the Escherichia coli strain BL21(DE3) Is an improvement to each of the basic parts. This system allows us to control when the gene is being expressed. This is a great advantage to us as large batch culture verexpression is very stressful on the cells. If we did not use an inducible promoter on our genes we would see significantly less pigment than our literature would lead us to expect. This is due to the fact that the genes we are using have been optimized to produce the maximum amount of pigment possible, and this process on a single young cell would be very stressful and significantly increase the doubling time of the cells. We have induced the cells after they have replicated many times and matured, this allows us to have more cells with the construct that can produce more pigment faster.

 Anthocyanin Constructs


Anthocyanin composite part 1

BBa_K1281101
T7 Promoter Part BBa_I712074.
RBS Part BBa_B0034.
dfr Part BBa_K1497010
f3h Part BBa_K1497009
Terminator Part BBa_B0015.


Anthocyanin composite part 2

BBa_K1281102
T7 Promoter Part BBa_I712074.
RBS Part BBa_B0034.
ans Part BBa_K1497002
Terminator Part BBa_B0015.


Anthocyanin composite part 3

BBa_K1281103
T7 Promoter Part BBa_I712074.
RBS Part BBa_B0034.
3gt Part BBa_K1281003
yadH Part BBa_K1281002
Terminator Part BBa_B0015.

These composite parts come together to complete the pathway from Eriodictyol to Anthocyanin. These constructs are separated due to the size of the genes. It would be too much of a sstrain on the cell to have all of th genes in one or even two plasmids and this is why we needed three separate constructs. These constructs are each submitted





 Zeaxanthin Construct

Zeaxanthin composite part

BBa_K1281301
T7 Promoter Part BBa_I712074.
RBS Part BBa_B0034.
crtY Part BBa_I742154.
crtZ Part BBa_I742157.
Terminator Part BBa_B0015.

This construct is in the plasmid psB1C3, and will convert our initial molecule Lycopene into our final product, the pigment Zeaxanthin.



 Indigoidine Constructs

Indigoidine composite part 1

BBa_K1281201
T7 Promoter PartBBa_I712074.
RBS Part BBa_B0034.
indB Part BBa_K1281001
Terminator Part BBa_B0015.


Indigoidine composite part 2

BBa_K1281202
T7 Promoter PartBBa_I712074.
RBS Part BBa_B0034.
indC Part BBa_K1152013
Terminator Part BBa_B0015.

These two composite parts come together to convert our initial molecule Glutamine into Indigoidine. The indB has been shown to increase the yields of Indigoidine as well as it is our original basic part submissions. This construct is part BBa_K1281201 and part BBa_K1281202. We had to split our genes into tow separate composite part submissions as the size of each was too large to allow for them to be in one plasmid.




 Melanin Construct

Melanin composite part

The parts used in our Melanin construct are as follows:
T7 Promoter PartBBa_I712074.
RBS Part BBa_B0034.
MelA Part BBA_K274001.
Terminator Part BBa_B0015.

This construct allows us to make Melanin out of L-Tyrosine.