Difference between revisions of "Team:ICT-Mumbai/Parts"

Line 58: Line 58:
 
}
 
}
  
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<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">
 
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Our project involves metabolism of ammonia by Escherichia coli to produce a blue-coloured dye,
 
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indigoidine. We had to choose between using a constitutive and an inducible promoter to drive
 
−
expression of the genes that we wish to express in E. coli.</p>
 
  
−
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">Expression from inducible promoters requires addition of an inducing molecule. However, as it would be
 
−
cumbersome and tedious to add the inducer to the device that will house the engineered E. coli, and as
 
−
it would also contribute to the cost, we decided to use a constitutive promoter to drive gene expression.</p>
 
−
 
−
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">We reasoned that the constitutive promoter of choice should have the following two properties: (1) it
 
−
should not be a very strong promoter, so as to not lead to any toxicity to the cell, and (2) it should be
 
−
active in low-nutrient conditions. Based on these two considerations, the commonly used glycolytic
 
−
promoters were ruled out as possible choices.</p>
 
−
 
−
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">The ychH promoter has been described in literature to be active under low glucose conditions (Ref. 1).
 
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Moreover, it is not a very strong promoter, compared to those frequently employed to express
 
−
recombinant proteins in E. coli (Ref. 2). Therefore, the ychH promoter became our promoter of choice.</p>
 
  
  
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</div>
 
</div>
  
 +
<div class="w3-content w3-container w3-padding-64" id="about">
 +
<div>
  
 +
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">
 +
Our project involves metabolism of ammonia by Escherichia coli to produce a blue-coloured dye,
 +
indigoidine. We had to choose between using a constitutive and an inducible promoter to drive
 +
expression of the genes that we wish to express in E. coli.</p>
  
 +
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">Expression from inducible promoters requires addition of an inducing molecule. However, as it would be
 +
cumbersome and tedious to add the inducer to the device that will house the engineered E. coli, and as
 +
it would also contribute to the cost, we decided to use a constitutive promoter to drive gene expression.</p>
  
−
<!--<div class="column full_size">
+
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">We reasoned that the constitutive promoter of choice should have the following two properties: (1) it
 +
should not be a very strong promoter, so as to not lead to any toxicity to the cell, and (2) it should be
 +
active in low-nutrient conditions. Based on these two considerations, the commonly used glycolytic
 +
promoters were ruled out as possible choices.</p>
  
 +
<p style="height:70 px; font-family:'Lato'; font-size:20px; colour:lightgrey;">The ychH promoter has been described in literature to be active under low glucose conditions (Ref. 1).
 +
Moreover, it is not a very strong promoter, compared to those frequently employed to express
 +
recombinant proteins in E. coli (Ref. 2). Therefore, the ychH promoter became our promoter of choice.</p>
 +
 +
 +
 +
 +
<!--<div class="column full_size">
 
<h1>Parts</h1>
 
<h1>Parts</h1>
  

Revision as of 03:06, 1 November 2017

ICT-Mumbai 2017

Our project involves metabolism of ammonia by Escherichia coli to produce a blue-coloured dye, indigoidine. We had to choose between using a constitutive and an inducible promoter to drive expression of the genes that we wish to express in E. coli.

Expression from inducible promoters requires addition of an inducing molecule. However, as it would be cumbersome and tedious to add the inducer to the device that will house the engineered E. coli, and as it would also contribute to the cost, we decided to use a constitutive promoter to drive gene expression.

We reasoned that the constitutive promoter of choice should have the following two properties: (1) it should not be a very strong promoter, so as to not lead to any toxicity to the cell, and (2) it should be active in low-nutrient conditions. Based on these two considerations, the commonly used glycolytic promoters were ruled out as possible choices.

The ychH promoter has been described in literature to be active under low glucose conditions (Ref. 1). Moreover, it is not a very strong promoter, compared to those frequently employed to express recombinant proteins in E. coli (Ref. 2). Therefore, the ychH promoter became our promoter of choice.