Coding
Part:BBa_K4165256
Designed by: Esraa Elmligy Group: iGEM22_CU_Egypt (2022-10-08)
GST-Coh2-G4S-WWW
Long description
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 85
WetLab Results
Transformation of GST COH WWW in BL-21 using pGS-21a vector
Figure 1. Transformed plate of GST COH (L) WWW + pGS-21a
Transformation of GST COH WWW in DH-5 alpha using pJET vector
Figure 2. Transformed plate of GST COH (L) WWW + pJET
Comparison between chemical lysis and sonication for GST COH WWW
Figure 3. This graph shows a significant difference between chemical lysis and sonication for GST COH WWW, after we had the results we optimized our protocol to use chemical lysis for GST COH WWW
SDS PAGE for induced and non induced samples of GST COH WWW
Figure 4. This figure shows the comparison between induced and non induced samples of GST COH WWW, where well no.1 is the induced sample while well no.5 is the non induced sample showing that our protein is induced effectively owing to our right choice of IPTC, time interval and concentration
Pull down assay of His Trim21 (L) against GST COH WWW and GST COH TD28Rev
Figure 5. This graph shows the comparison of pull down assay between His trim21(L) DOC with GST COH WWW and GST COH TD28Rev, showing that the interaction between His Trim21 (L) DOC and GST COH WWW is better than that of His Trim21 (L) DOC and GST COH TD28Rev as the concentration of the elution of His Trim21 (L) DOC with GST COH WWW is more than that of His Trim21 (L) DOC with GST COH TD28Rev
Pull down assay of Tau aggregates against GST COH WWW and GST COH TD28Rev
Figure 6. This graph shows the comparison of pull down assay between Tau aggregates with GST COH WWW and GST COH TD28Rev, showing that the interaction between Tau aggregates with GST COH WWW is better than that of Tau aggregates with GST COH TD28Rev as the concentration of elution of Tau aggregates with GST COH WWW is more than that of Tau aggregates with GST COH TD28Rev, illustrating that WWW could be a better candidate for binding to the Tau aggregates
BCA assay results of GST COH WWW
Figure 7. This graph illustrates the results of BCA assay for GST COH WWW showing that our protein concentration is expected to be 0.719466667
[edit]
Categories
Parameters
None |