Difference between revisions of "Part:BBa K4165169"
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                                 Figure 4. Transformed plate of His COH + pJET  
 
                                 Figure 4. Transformed plate of His COH + pJET  
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<p style=" font-weight: bold; font-size:14px;"> Comparison between chemical lysis and sonication for His COH </p>
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<p><img src="https://static.igem.wiki/teams/4165/wiki/data-analysis/sonication-or-chemical/sonication-or-chemical/his-coh.jpg" style="margin-left:200px;" alt="" width="500" /></p>
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Figure 5. This graph shows the difference between chemical lysis and sonication for His COH, after we had the results we optimized our protocol to use sonication for His COH
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<!-- Uncomment this to enable Functional Parameter display  
 
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Revision as of 14:25, 11 October 2022


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

Transformation of His COH in BL-21 using pGS-21a vector 

                                 Figure 3. Transformed plate of His Coh + pGS-21a

Transformation of His COH in DH-5 alpha using pJET vector 

                               Figure 4. Transformed plate of His COH + pJET

Comparison between chemical lysis and sonication for His COH

Figure 5. This graph shows the difference between chemical lysis and sonication for His COH, after we had the results we optimized our protocol to use sonication for His COH