Difference between revisions of "Part:BBa K2933218"
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This composite part is made up with seven basic parts, the His tag, T7 promoter, RBS and our target protein PST-1. It encodes a protein which is PST-1 fused with His tag. The fusion protein is about 26.2 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of PST-1. It is convenient for us to purify our target protein.<br> | This composite part is made up with seven basic parts, the His tag, T7 promoter, RBS and our target protein PST-1. It encodes a protein which is PST-1 fused with His tag. The fusion protein is about 26.2 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of PST-1. It is convenient for us to purify our target protein.<br> | ||
===Molecular cloning=== | ===Molecular cloning=== | ||
− | First, we used the vector pGEX- | + | First, we used the vector pGEX-28a to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.<br> |
<p style="text-align: center;"> | <p style="text-align: center;"> | ||
[[File:PST-1-PCR1.png]]<br> | [[File:PST-1-PCR1.png]]<br> | ||
'''Figure 1.''' The PCR result of PST-1.<br> | '''Figure 1.''' The PCR result of PST-1.<br> | ||
</p> | </p> |
Revision as of 13:06, 23 September 2019
T7 promoter+RBS b+Linker h+His+Linker f+PST-1+T7 terminator
This part consists of T7 promoter, RBS and protein coding sequence(His+Linker f+PST-1),and the biological module can be built into E.coli for protein expression.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 47
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 169
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 47
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 47
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology=
This composite part is made up with seven basic parts, the His tag, T7 promoter, RBS and our target protein PST-1. It encodes a protein which is PST-1 fused with His tag. The fusion protein is about 26.2 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of PST-1. It is convenient for us to purify our target protein.
Molecular cloning
First, we used the vector pGEX-28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.