Difference between revisions of "Part:BBa K2933169"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | This composite part is made up with | + | This composite part is made up with four basic parts(Tac promoter,RBS a , Linker g , GST+Linker e+MUS-2).It encodes a protein which is MUS-2 fused with GST tag. The fusion protein is about 54.3 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MUS-2 and combine the two parts with the cutting site of Prescission Protease. The fusion protein can be cut off at the cutting site by Prescission Protease. It is convenient for us to purify our target protein.<br> |
===Molecular cloning=== | ===Molecular cloning=== |
Revision as of 08:27, 24 September 2019
Tac promoter+RBS a+Linker g+GST+Linker e+MUS-2
This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+MUS-2),and the biological module can be built into E.coli for protein expression.
Sequence and Features
- 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 181
Usage and Biology
This composite part is made up with four basic parts(Tac promoter,RBS a , Linker g , GST+Linker e+MUS-2).It encodes a protein which is MUS-2 fused with GST tag. The fusion protein is about 54.3 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MUS-2 and combine the two parts with the cutting site of Prescission Protease. The fusion protein can be cut off at the cutting site by Prescission Protease. It is convenient for us to purify our target protein.
Molecular cloning
We used the vector pGEX-6p-1 to construct our expression plasmid.
Figure 1. Left: The PCR result of MUS-2. Right: The verification results by enzyme digestion.
After verification, it was determined that the construction is successful.