Difference between revisions of "Part:BBa K2933209"
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[[File:PEDO-1-PCR.png|500px]]<br> | [[File:PEDO-1-PCR.png|500px]]<br> | ||
− | '''Figure 1.''' | + | '''Figure 1.''' The PCR result of PEDO-1. <br> |
</p> | </p> |
Revision as of 12:22, 23 September 2019
T7 promoter+RBS b+Linker h+His+Linker f+PEDO-1+T7 terminator
This part consists of T7 promoter, RBS and protein coding sequence(His+Linker f+PEDO-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
Illegal PstI site found at 741
Illegal PstI site found at 1027 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 169
Illegal PstI site found at 741
Illegal PstI site found at 1027 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 47
Illegal PstI site found at 741
Illegal PstI site found at 1027 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 47
Illegal PstI site found at 741
Illegal PstI site found at 1027 - 1000COMPATIBLE WITH RFC[1000]
Usage and Biology=
This composite part is made up with three basic parts, the His tag, T7 promoter, RBS and our target protein PEDO-1. It encodes a protein which is PEDO-1 fused with His tag. The fusion protein is about 32.2 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of PEDO-1 . It is convenient for us to purify our target protein.
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.