Difference between revisions of "Part:BBa K2933217"
(→Usage and Biology=) |
|||
| Line 19: | Line 19: | ||
<!-- --> | <!-- --> | ||
==Usage and Biology=== | ==Usage and Biology=== | ||
| − | This composite part is made up with | + | This composite part is made up with seven 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.<br> |
===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. | ||
Revision as of 13:00, 23 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
Assembly Compatibility:
- 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 seven 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. The result of PCR