Part:BBa_K676013
Mu/Spy Ligation
A device to detect the level of stress inside a cell due to the presence of a GBS on the plasmid.
Usage and Biology
The Mu GBS which was ligated into the BBa_K239009 SPY device shows a higher expression of GFP in vivo compared to the SPY device without the Mu GBS . The data was plotted into a graph and presented as a characterisation of device in our project. The higher expression level may due to the fact that negative supercoiling in the DNA promote easier transition from the close promoter complex to open promoter complex for higher rate of transcription. The higher level of expression means that the introduction of such GBS into DNA vaccine or gene therapy plasmid will be able to trigger a better immune response for a fully developed protection against the target infection .Although this is only in theory , further studies will be required to vertify it.
The higher level of expression can be use in making a more sensitive SPY device in order to detect shear stress level of the cells during fermentation as well as in downstream processing.
As with the BBa_K676001( Mu GBS ) , we also observed a slightly higher supercoiling efficiency ( about 10% higher compare to the controls) and better uniformity (smaller range of supercoiled topoisomers) in the supercoiled plasmids (Mu GBS with SPY device) .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 191
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1203
| n/a | Mu/Spy Ligation |