Difference between revisions of "Part:BBa K4165072"
Line 48: Line 48:
 
     the variation of mRNA and protein concentrations with time compared with the wet lab results (2mM IPTG concentration)
 
     the variation of mRNA and protein concentrations with time compared with the wet lab results (2mM IPTG concentration)
 
===WetaLab Results===
 
===WetaLab Results===
 +
<p style=" font-weight: bold; font-size:14px;"> Transformation of GST DOC in BL-21 using pGS-21a vector </p>
 
<html>
 
<html>
 
<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/gst-doc-pgs.jpg" style="margin-left:200px;" alt="" width="500" /></p>
 
<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/gst-doc-pgs.jpg" style="margin-left:200px;" alt="" width="500" /></p>
 
</html>
 
</html>
 
                                   Figure 4. Transformed plate of GST Doc + pGS-21a  
 
                                   Figure 4. Transformed plate of GST Doc + pGS-21a  
 +
<p style=" font-weight: bold; font-size:14px;"> Transformation of GST DOC in DH-5 alpha using pJET vector </p>
 +
<html>
 +
<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/gst-doc-pjet.jpg" style="margin-left:200px;" alt="" width="500" /></p>
 +
</html>
 +
                                  Figure 5. Transformed plate of GST DOC + pJET
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 11:58, 11 October 2022


T7 promoter-GST-DocS-T7 terminator

This composite part is composed of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), RBS 4 (BBa_K4165264), GST-tag (BBa_K4165070), DocS (BBa_K3396000), and T7 terminator (BBa_K731721).

Source

Synthesized

Usage and Biology

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

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

Dry Lab

Modeling

Model “peptide (GST_DocS) modeled by Rosettafold full_model_407305_3” Ranked fourth model from our 4 ones with score 4 out of 6 and values of C Beta deviation = 0 , clash score = 182.78 , molprobitey = 2.72 , Ramachandran favored = 98.25 , Ramachandran outlier = 0.7 , Q Mean_4 = 0.75 , Q Mean_6 = 1.02 


                               Figure 1. The 3D structure of GST-DOCs 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 (1mM IPTG concentration)



              Figure 3. 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 (2mM IPTG concentration)

WetaLab Results

Transformation of GST DOC in BL-21 using pGS-21a vector 

                                  Figure 4. Transformed plate of GST Doc + pGS-21a

Transformation of GST DOC in DH-5 alpha using pJET vector 

                                  Figure 5. Transformed plate of GST DOC + pJET