Difference between revisions of "Part:BBa K2933217"
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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. | ||
− | [[File:VIM-66- | + | [[File:VIM-66-PCR1.jpeg|600px|center|]] |
<p style="text-align: center;"> | <p style="text-align: center;"> | ||
'''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 | '''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 |
Revision as of 14:34, 21 September 2019
T7 promoter+RBS b+Linker h+His+Linker f+VIM-66+T7 terminator
This part consists of T7 promoter, RBS and protein coding sequence(His+Linker f+VIM-66),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
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 942
- 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 three basic parts, the His tag, T7 promoter, RBS and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of VIM-66 . 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