Difference between revisions of "Part:BBa K2933229"
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===Usage and Biology=== | ===Usage and Biology=== | ||
This composite part is made up with five basic parts, the RBSa, the linker g and the GST tag, the cutting site of Prescission Protease and our target protein PEDO-1. It encodes a protein which is PEDO-1 fused with GST tag. The fusion protein is about 58.2 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of PEDO-1 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> | This composite part is made up with five basic parts, the RBSa, the linker g and the GST tag, the cutting site of Prescission Protease and our target protein PEDO-1. It encodes a protein which is PEDO-1 fused with GST tag. The fusion protein is about 58.2 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of PEDO-1 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> | ||
| + | ===References=== | ||
| + | [1]Dereje Dadi Gudeta, Valeria Bortolaia,The Soil Microbiota Harbors a Diversity of Carbapenem-Hydrolyzing β-Lactamases of Potential Clinical Relevance[J]，Antimicrobial Agents and chemotherapy,January 2016<br> | ||
===Molecular cloning=== | ===Molecular cloning=== | ||
Latest revision as of 09:15, 24 September 2019
RBS a+Linker g+GST+Linker e+PEDO-1
This part consists of RBS a, protein coding sequence(GST+Linker e+PEDO-1), the RBS and the protein coding sequence can be connected by linker g. The biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1263
Illegal PstI site found at 1549 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1263
Illegal PstI site found at 1549 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1263
Illegal PstI site found at 1549 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1263
Illegal PstI site found at 1549 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 113
Usage and Biology
This composite part is made up with five basic parts, the RBSa, the linker g and the GST tag, the cutting site of Prescission Protease and our target protein PEDO-1. It encodes a protein which is PEDO-1 fused with GST tag. The fusion protein is about 58.2 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of PEDO-1 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.
References
[1]Dereje Dadi Gudeta, Valeria Bortolaia,The Soil Microbiota Harbors a Diversity of Carbapenem-Hydrolyzing β-Lactamases of Potential Clinical Relevance[J]，Antimicrobial Agents and chemotherapy,January 2016
Molecular cloning
First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.
Figure 1. Left: The PCR result of PEDO-1. Right: The verification results by enzyme digestion.