Difference between revisions of "Part:BBa K4165167"
Line 42: Line 42:
  
 
===WetLab Results===
 
===WetLab Results===
 +
<p style=" font-weight: bold; font-size:14px;"> Transformation of His 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/his-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/his-doc-pgs.jpg" style="margin-left:200px;" alt="" width="500" /></p>
 
</html>
 
</html>
 
                                     Figure 3. Transformed plate of His Doc + pGS-21a  
 
                                     Figure 3. Transformed plate of His Doc + pGS-21a  
 +
<p style=" font-weight: bold; font-size:14px;"> Transformation of His Doc in DH-5 alpha using pJET vector </p>
 +
<html>
 +
<p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/his-doc-pjet.jpg" style="margin-left:200px;" alt="" width="500" /></p>
 +
</html>
 +
                                    Figure 4. Transformed plate of His Doc + pJET
 +
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K4165167 parameters</partinfo>
 
<partinfo>BBa_K4165167 parameters</partinfo>
 
<!-- -->
 
<!-- -->

Revision as of 12:11, 11 October 2022


His - DocS

This composite part consists of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), RBS 4 (BBa_K4165264), 6x His-tag (BBa_K4165020), DocS (BBa_K3396000) and T7 terminator (BBa_K731721).

Source

Synthesized

Usage and Biology

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

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

Dry Lab

Modeling

Model “peptide (His_DocS) modeled by TRrosetta model1” Ranked Third model from our 4 ones with score 5 out of 6 and values of C Beta deviasion = 0 , clash score = 0 , molprobitey = 0.77 , Ramachandran favored = 96 , Ramachandran outlier = 0 , Q Mean_4 = -0.29 , Q Mean_6 = -0.27 



                          Figure 1. The 3D structure of His 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.

WetLab Results

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

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

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

                                    Figure 4. Transformed plate of His Doc + pJET