Part:BBa_K4165232
(GST) COH2-linker(G4S)-TD28REV
This biobrick consists of T7 promotor (BBa_K3633015), RBS (BBa_K4165016), GST (BBa_K4165070), COH2 (BBa_K4165003), G4S linker (BBa_K4165068), TD28REV (BBa_K4165006), Terminator (BBa_K731721), The GST tag was attached to the (COH2-Linker-TD28REV) coding sequence to serve in the purification using Glutathione resin column.
Usage and Biology
The main function is the binding of the tau-binding peptide with the Cohasin protein. It has been used the project to target and degrade both tau and Aβ proteins which are both considered the main causes of Alzheimer’s Disease pathogenesis.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 45
Illegal SapI.rc site found at 189
Modeling
For 2 Models with a score of 5 out of 6 and among 11 Models, we choose our best-ranked model according to its score of 5 out of 6 Q Mean_4 value. Our model values :
| cbeta_deviations | clashscore | molprobity | ramachandran_favored | ramachandran_outliers | Qmean_4 | Qmean_6 |
|---|---|---|---|---|---|---|
| 1 | 2.74 | 1.22 | 97.11 | 0.52 | 1.96 | 1.88 |
Figure 1. The 3D structure of the COH-linker-TD28rev model Displayed on Pymol.
Mathematical modeling
Transcription rate and translation rate under T7 promotor
the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in the code section) beside with the estimated results from the wet lab.
Figure 2. this figure shows the results from the transcription and translation code showing the
variation of mRNA and protein concentrations with time compared with the wet lab results.
WetLab Results
Comparison between the mathematical model of dry lab and the wey lab results of GST COH TD28 Rev using chemical lysis
Figure 3. This graph shows that the mathematical model of the dry lab and the wet lab results of GST COH
TD28Rev using chemical lysis doesn't correlate with each other as there is a great difference between the
two results.
Comparison between the mathematical model of dry lab and the results of wet lab of GST COH TD28Rev using sonicator
Figure 4. This graph shows that there is a correlation between the mathematical model of the dry lab and the
wet lab results of GST COH TD28Rev using sonicator
| None |