Difference between revisions of "Part:BBa K2933175"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | <br> | + | This composite part is made up with three basic parts(Tac promoter，RBS a and Linker g)and a composite part(GST+Linker e+VIM-66).It encodes a protein which is VIM-66 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 VIM-66 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> |
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===Molecular cloning=== | ===Molecular cloning=== | ||
We insert VIM-66 gene into the standard vector then transfer it into E.coli. | We insert VIM-66 gene into the standard vector then transfer it into E.coli. | ||
Revision as of 14:29, 21 September 2019
Tac promoter+RBS a+Linker g+GST+Linker e+VIM-66
This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+VIM-66),and the biological module can be built into E.coli for protein expression.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1532
- 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 three basic parts(Tac promoter，RBS a and Linker g)and a composite part(GST+Linker e+VIM-66).It encodes a protein which is VIM-66 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 VIM-66 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 insert VIM-66 gene into the standard vector then transfer it into E.coli.
Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification