Difference between revisions of "Part:BBa K2933155"
Line 17: | Line 17: | ||
<partinfo>BBa_K2933155 parameters</partinfo> | <partinfo>BBa_K2933155 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
+ | ===Usage and Biology=== | ||
+ | This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site 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. 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. | ||
+ | [[File:VIM-66-PCR.jpeg|600px|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 |
Revision as of 07:56, 22 September 2019
His+Linker f+VIM-66
This part encodes the fusion protein of His tag and VIM-66 to promote the expression and purification of target protein(VIM-66).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 824
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site 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. 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