Difference between revisions of "Part:BBa K4165171"
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===WetLab Results=== | ===WetLab Results=== | ||
+ | <p style=" font-weight: bold; font-size:14px;"> Transfotmation of GST COH 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-coh.jpg" style="margin-left:200px;" alt="" width="500" /></p> | <p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/gst-coh.jpg" style="margin-left:200px;" alt="" width="500" /></p> | ||
</html> | </html> | ||
− | Figure 2. Transformed plate of GST COH | + | Figure 2. Transformed plate of GST COH + pGS-21a |
+ | <p style=" font-weight: bold; font-size:14px;"> Transformation of GST COH in DH-5 alpha using pJET vector </p> | ||
+ | <html> | ||
+ | <p><img src="https://static.igem.wiki/teams/4165/wiki/parts-registry/wetlab-results/gst-coh-pjet.jpg" style="margin-left:200px;" alt="" width="500" /></p> | ||
+ | </html> | ||
+ | Figure 3. Transformed plate of GST COH + pJET | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K4165171 parameters</partinfo> | <partinfo>BBa_K4165171 parameters</partinfo> | ||
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Revision as of 11:09, 11 October 2022
Biobrick GST - CoH2
This composite part consists of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), pGS-21a RBS (BBa_K4165016), GST tag (BBa_K4165070), 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 GST tag for the purification of the COH.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1172
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 45
Illegal SapI.rc site found at 189
Dry lab
Modeling
Model “peptide (GST_Coh2) modeled by Alphafold GST_Coh2_f66fb_unrelaxed_rank_3_model_2” Ranked first model from our 4 ones with score 5 out of 6 and values of C Beta deviation = 0 , clash score = 13.59 , molprobitey = 1.64 , Ramachandran favored = 98.31 , Ramachandran outlier = 0.56 , Q Mean_4 = 0.643, Q Mean_6 = 0.914
Figure 1. The 3D structure of 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 1. 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
Transfotmation of GST COH in BL-21 using pGS-21a vector
Figure 2. Transformed plate of GST COH + pGS-21a
Transformation of GST COH in DH-5 alpha using pJET vector
Figure 3. Transformed plate of GST COH + pJET