Difference between revisions of "Part:BBa K3419004"
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− | <td> | + | <td>bas<sub>T</sub></td> |
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<td>23.22 micromolar/minute</td> | <td>23.22 micromolar/minute</td> | ||
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<td>degradation rate of mCardinal</td> | <td>degradation rate of mCardinal</td> | ||
− | <td>0.575 min | + | <td>0.575 min<sup>-1</sup></td> |
<td>[5] </td> | <td>[5] </td> | ||
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Revision as of 21:28, 17 October 2020
mCardinal with Strong Promoter
Usage and Biology
mCardinal is a red fluorescent protein that absorbs light of peak wavelength 604 nm and emits light of peak wavelength 659 nm, derived from Entacmaea quadricolor. This BioBrick part consists of the mCardinal coding gene [1] with stop codon TAA. The sequence was reverse translated with codon optimization for E. Coli. It is driven by a constitutive promoter (BBa_J23100), has a ribosome binding site (BBa_J61100), and a terminator (BBa_B0015). The constitutive promoter was chosen to allow for maximum and constant mCardinal expression. After reviewing several sources, we were able to find that mCardinal could realistically be produced by E. coli cells. Chu et al (2014) was able to study the mCardinal fluorescent protein after purifying it from cultured E. coli cells that were grown to produce the protein [2].
In order to determine the location of the E. coli in the body, our system utilized a fluorescent reporter that can be detected by our sensing device, Trichoscan. We chose to use a red fluorescent protein due to the ease of detection; red light can best penetrate layers of the skin [3], and mCardinal is a relatively bright red fluorescent protein. Thus, the Trichoscan device can determine the location of the bacteria.
Modeling
In MATLAB, we modeled the output of the mCardinal protein using the following differential equation, and we solved it using MATLAB’s ode45 function. The equation represents the production of mCardinal protein. The first term represents the basal expression under the constitutive promoter (BBa_J23100), while the second represents protein degradation.
The MATLAB scripts corresponding to the mCardinal modeling can be downloaded here:
File:ModelingCodeCornelliGEM2020.zip
Variable | Parameter | Value | Source |
---|---|---|---|
[mCardinal] | Concentration of mCardinal | -- | -- |
basT | Basal transcription rate | 23.22 micromolar/minute | [4] |
dC | degradation rate of mCardinal | 0.575 min-1 | [5] |