Adityakamat (Talk | contribs) |
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+ | <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> | ||
+ | |||
+ | <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> | ||
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Revision as of 02:57, 1 November 2017