Difference between revisions of "Team:Hong Kong UCCKE/Basic Part"
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| − | <h3 class="sectiontitle"> | + | <h3 class="sectiontitle">BBa_K2197300 |
| + | </h3> | ||
| + | <h4 class="sectiontitle">Purpose</h4> | ||
| + | |||
<div class="divider"></div> | <div class="divider"></div> | ||
| − | <p style="text-align:left !important;"> | + | <p style="text-align:left !important;">Engineered e.coli encodes part BBa_K2197300. By adding blood samples to culture medium which engineered e.coli is cultured, e.coli expresses different level of GFP. By analysing the GFP level with HPLC, the uric acid concentration of the sample can be estimated. This part ensures a rapid detection of uric acid concentration thus gout. |
| − | <p style="text-align:left !important;"> | + | </p> |
| − | + | <h4 class="sectiontitle">Design</h4> | |
| − | <h3 class="sectiontitle" style="clear:both;"> | + | <p style="text-align:left !important;">Part BBa_K2197300 can be divided into two sessions, the promoter and the GFP expression. The promoter is designed to be sensitive to the concentration of uric acid. This promoter control the expression of GFP that is downstream the promoter. The promoter session consists of a constitutive promoter J23100, a RBS B0034, a strong repressor KRAB-HucR, a double terminator B0015. HucR is itself a repressor. Its repressing ability is enhanced by KRAB. The resulting repressor is a chimeric mammalian urate-dependent transsilencer (mUTS). hucO is an operative site for mUTS to bind. When mUTS is binded to hucO, the expression of downstream gene is restricted according concentration of substrates. The presence of uric acid limits the binding of mUTS to hucO. The limitation varies as the concentration of uric acid. Downstream of the promoter session is a constitutive promoter J23106, a RBS B0034, a GFP gene E0040 and a terminator B1002. As a result mUTS binds to hucO and GFP is not expressed when uric acid is absent or at very low concentration. Alternatively, the complex detaches from hucO and GFP is expressed according to the concentration of uric acid. The promoter control the expression of GFP. Engineered e.coli encodes part BBa_K2197300. |
| + | </p> | ||
| + | <h4 class="sectiontitle"> Calibration</h4> | ||
| + | |||
| + | <p style="text-align:left !important;">graphs | ||
| + | GFP expression regulated by a concentration-sensitive-promoter (see link) | ||
| + | http://www.nature.com/nbt/journal/v28/n4/full/nbt.1617.html | ||
| + | </p> | ||
| + | |||
| + | |||
| + | <h3 class="sectiontitle" style="clear:both;"> | ||
| + | BBa_K2197301 | ||
| + | </h3> | ||
<div class="divider"></div> | <div class="divider"></div> | ||
| − | <p style="text-align:left !important;"> | + | <p style="text-align:left !important;">This is a composite part encoding hucO(BBa_K2197303) and GFP(E0040) gene. The circuit uses a bacterial transcriptional repressor (HucR) that binds a DNA sequence motif (hucO) in the absence of uric acid. When uric acid is present, HucR dissociates from hucO motif, thereby allowing expression of a downstream GFP gene. It is hoped that the expression of the downstream GFP gene is regulated by the concentration of uric acid in a proportional manner. |
| + | -- | ||
| + | http://www.nature.com/nbt/journal/v28/n4/full/nbt.1617.html | ||
| + | </p> | ||
</div> | </div> | ||
Revision as of 17:31, 13 October 2017
BBa_K2197300
Purpose
Engineered e.coli encodes part BBa_K2197300. By adding blood samples to culture medium which engineered e.coli is cultured, e.coli expresses different level of GFP. By analysing the GFP level with HPLC, the uric acid concentration of the sample can be estimated. This part ensures a rapid detection of uric acid concentration thus gout.
Design
Part BBa_K2197300 can be divided into two sessions, the promoter and the GFP expression. The promoter is designed to be sensitive to the concentration of uric acid. This promoter control the expression of GFP that is downstream the promoter. The promoter session consists of a constitutive promoter J23100, a RBS B0034, a strong repressor KRAB-HucR, a double terminator B0015. HucR is itself a repressor. Its repressing ability is enhanced by KRAB. The resulting repressor is a chimeric mammalian urate-dependent transsilencer (mUTS). hucO is an operative site for mUTS to bind. When mUTS is binded to hucO, the expression of downstream gene is restricted according concentration of substrates. The presence of uric acid limits the binding of mUTS to hucO. The limitation varies as the concentration of uric acid. Downstream of the promoter session is a constitutive promoter J23106, a RBS B0034, a GFP gene E0040 and a terminator B1002. As a result mUTS binds to hucO and GFP is not expressed when uric acid is absent or at very low concentration. Alternatively, the complex detaches from hucO and GFP is expressed according to the concentration of uric acid. The promoter control the expression of GFP. Engineered e.coli encodes part BBa_K2197300.
Calibration
graphs GFP expression regulated by a concentration-sensitive-promoter (see link) http://www.nature.com/nbt/journal/v28/n4/full/nbt.1617.html
BBa_K2197301
This is a composite part encoding hucO(BBa_K2197303) and GFP(E0040) gene. The circuit uses a bacterial transcriptional repressor (HucR) that binds a DNA sequence motif (hucO) in the absence of uric acid. When uric acid is present, HucR dissociates from hucO motif, thereby allowing expression of a downstream GFP gene. It is hoped that the expression of the downstream GFP gene is regulated by the concentration of uric acid in a proportional manner. -- http://www.nature.com/nbt/journal/v28/n4/full/nbt.1617.html
