Difference between revisions of "Part:BBa K676013"

Latest revision as of 22:17, 9 October 2011

Mu/Spy Ligation

A device to see the difference in the expression of GFP between the presence and absence of a gyrase binding site on a plasmid molecule.

Usage and Biology

The Mu phage GBS which was ligated to the Spy promoter device (Part:BBa_K239009) and it displayed a higher expression of GFP in vivo compared to the old Spy promoter device without the Mu phage GBS. The higher expression level may possibly be 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 therefore indicates that the introduction of such GBS into plasmids carrying DNA vaccine or therapeutic genes will be able to trigger a better immune response and hence induce a fully developed protection against the target infection. Although this is only a small experiment to confirm the higher expression, further studies are required to verify it.

The higher level of expression can be used to make a more sensitive Spy promoter device (shearometer) in order to detect lower shear stress levels in the cells during fermentation and downstream processing.

As with the Mu phage GBS (Part:BBa_K676001) ligated to the Spy promoter device, we also observed a slightly higher supercoiling efficiency (about 10%) and better uniformity (smaller range of supercoiled topoisomers) in the mini-prep plasmid sample when compared to the control plasmid (Spy promoter device with no GBS).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 191
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1203

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K676013 short</partinfo>
-A device to detects the level of stress due to a GBS being present on the plasmid.
+A device to see the difference in the expression of GFP between the presence and absence of a gyrase binding site on a plasmid molecule.
-<!-- Add more about the biology of this part here
 ===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 Mu phage GBS which was ligated to the Spy promoter device ([[Part:BBa_K239009]]) and it displayed a higher expression of GFP in vivo compared to the old Spy promoter device without the Mu phage GBS. The higher expression level may possibly be 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 therefore indicates that the introduction of such GBS into plasmids carrying DNA vaccine or therapeutic genes will be able to trigger a better immune response and hence induce a fully developed protection against the target infection. Although this is only a small experiment to confirm the higher expression, further studies are required to verify it.
-The higher level of expression can be use in making a more sensitive SPY device in order to detect shear stress during fermentation as well as in downstream processing.
+The higher level of expression can be used to make a more sensitive Spy promoter device (shearometer) in order to detect lower shear stress levels in the cells during fermentation and downstream processing.
+As with the Mu phage GBS ([[Part:BBa_K676001]]) ligated to the Spy promoter device, we also observed a slightly higher supercoiling efficiency (about 10%) and better uniformity (smaller range of supercoiled topoisomers) in the mini-prep plasmid sample when compared to the control plasmid  (Spy promoter device with no GBS).
+[[Image:Improving_BBa_K239009_into_BBa_K676013.jpg]]
+[[Image:Improving_Flour_BBa_K239009_into_BBa_K676013.jpg]]
-As with the BBa_K676001( Mu GBS ) , we also observed a better uniformity in the supercoiled plasmids ( Mu GBS with SPY device) . Although the extent of supercoiling in the plasmid with and without the GBS only showed a little difference but the GBS provided us with a more uniformly supercoiled plasmid products.
 <!-- -->