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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE 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