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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 539
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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
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