Part:BBa_K4165169

Biobrick His - CoH2

This composite part consists of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), RBS 4 (BBa_K4165264), 6x His-tag (BBa_K4165020), CoH2 (BBa_K4165003) and T7 terminator (BBa_K731721).

Source

Synthesized

Usage and Biology

COH protein is a part of the Snitch system, which binds to the His tag for the purification of the COH.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 539
  
• 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
  

Dry Lab

Modeling

Model “peptide (His_Coh2) modeled by TRrosetta model1” Ranked second model from our 4 ones with score 5 out of 6 and values of C Beta deviasion = 0 , clash score = 3.23 , molpro0bitey = 1.59 , Ramachandran favored = 92.31 , Ramachandran outlier = 1.4 , Q Mean_4 = 0.835 , Q Mean_6 = 0.95

                              Figure 1. The 3D structure of His-COH model visualized by 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 results of wet lab of His COH using sonicator

Comparison between the mathematical model of dry lab and the wet lab results of His COH using chemical lysis

Figure 4. This graph shows that the mathematical model of dry lab and the wet lab results of His COH using chemical lysis don't correlate with each other as there is a significant difference between the results