Difference between revisions of "Test XOR Hybrid Promoters"

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This is a rough idea of how to test the four hybrid promoters for the XOR gate.
+
This are our ideas of how to test the four hybrid promoters for the XOR gate.
  
  
'''Test pLuxR/cI'''
+
[[Test pLuxR/cI]]
  
luxR/CI hybrid promoter is constitutively off. Ligate with GFP on the 3’ end. If luxR is present first then add AI-1, the bacteria should glow green. Then add CI, the bacteria should not glow.
 
  
Some promoter→LuxR+t+t+pLuxR/cI→GFP does not glows
 
                                                     
 
                                                         
 
add AI-1
 
  
Some promoter→LuxR+t+t+pLuxR/cI→GFP glows
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[[Test pMnt/Lac]]
  
add AI-1
 
  
Some promoter→LuxR+cI+t+t+pLuxR/cI→GFP does not glow
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[[Test pLsrA/cI]]
  
 
{| border="1" cellpadding="2"
 
!width="30"|LuxR
 
!width="30"|cI
 
!width="30"|AI-1 + GFP
 
!width="30"|AI-1 - GFP
 
|-
 
| 0 || 0 || 0 || 0
 
|-
 
| 1 || 0 || 1 || 0
 
|-
 
| 0 || 1 || 0 || 0
 
|-
 
| 1 || 1 || 0 || 0
 
|}
 
  
  
 +
[[Test pLasR/cI]]
  
  
  
'''Test pMnt/Lac'''
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[[Test Mnt/Tet promoter]]
  
pMnt/Lac is constitutively on but is repressed by the LacI protein. The promoter is also repressed by the Mnt protein. The addition of IPTG should repress the LacI repressor and glow in the absence of the Mnt protein.
 
  
Some promoter→LacI+t+t+pLac/Mnt→GFP    does not glow
 
 
Some promoter→Mnt+t+t+pLac/Mnt→GFP    does not glow
 
 
                                                       
 
add IPTG
 
 
Some promoter→LacI+t+t+pLac/Mnt→GFP    glows
 
  
 +
'''Overlapping Parts And intermediates'''
  
  
 
{| border="1" cellpadding="2"
 
{| border="1" cellpadding="2"
!width="30"|LacI
+
!width="30"|LuxR/cI
!width="30"|Mnt
+
!width="30"|Lac/Mnt
!width="30"|IPTG + GFP
+
!width="30"|LsrR/cI
!width="30"|IPTG - GFP
+
!width="30"|LasR/cI
 +
!width="30"|Mnt/Tet
 
|-
 
|-
| 0 || 0 || 1 || 1
+
| RBS+cI(+TT) || || RBS+cI(+TT) ||  ||  
 
|-
 
|-
| 0 || 1 || 0 || 0
+
| || RBS+Mnt(+TT) || || RBS+Mnt(+TT) ||  
 
|-
 
|-
| 1 || 0 || 1 || 0
+
| || || ||  ||
 
|-
 
|-
| 1 || 1 || 0 || 0
+
| || ||  || ||
 
|}
 
|}
 
 
 
 
'''Test pLsrR/cI'''
 
 
pLsrR/cI is constitutively on and is repressed by LsrR. The addition of AI-2, which becomes phosphorylated by LsrK, represses LsrR causing the promoter to turn on. The addition of cI to should repress the promoter causing it to turn off.
 
 
Some promoter→LsrK+LsrR+t+t+pLsrR/cI→GFP  does not glow
 
 
 
add AI-2             
 
                                               
 
Some promoter→LsrK+LsrR+t+t+pLsrR/cI→GFP glows
 
 
 
add AI-2
 
                                                             
 
Some promoter→LsrK+LsrR+cI+t+t+pLsrR/cI→GFP does not glows
 
 
{| border="1" cellpadding="2"
 
!width="30"|LsrR
 
!width="30"|cI
 
!width="30"|AI-1 + GFP
 
!width="30"|AI-1 - GFP
 
|-
 
| 0 || 0 || 1 || 1
 
|-
 
| 1 || 0 || 1 || 0
 
|-
 
| 0 || 1 || 0 || 0
 
|-
 
| 1 || 1 || 0 || 0
 
|}
 
 
 
 
 
 
'''Test pMnt/Tet'''
 
 
The pMnt/TetR promoter is constitutively on and is repressed by the TetR protein. The addition of aTc should repress the TetR causing it to glow. The addition of the Mnt protein should repress the promoter causing it not glow.
 
 
 
 
{| border="1" cellpadding="2"
 
!width="30"|TetR
 
!width="30"|Mnt
 
!width="30"|aTc + GFP
 
!width="30"|aTc - GFP
 
|-
 
| 0 || 0 || 1 || 1
 
|-
 
| 0 || 1 || 0 || 0
 
|-
 
| 1 || 0 || 1 || 0
 
|-
 
| 1 || 1 || 0 || 0
 
|}
 
 
 
Test Tet repression:
 
 
Some promoter (pbad)→RBS+TetR+t+t+pMnt/Tet→RBS+GFP does not glows
 
 
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I13453 I13453]+RBS+[http://partsregistry.org/Part:BBa_C0040 C0040]+TT+[http://partsregistry.org/Part:BBa_K091105 K091105]+[http://partsregistry.org/Part:BBa_E0240 E0240]
 
 
 
add aTc
 
                                                               
 
Some promoter→TetR+t+t+pMnt/Tet→GFP glows
 
 
I13453+RBS+C0040+TT+K091105+E0240
 
 
 
Test Mnt repression:
 
 
Some promoter→Mnt+t+t+pMnt/Tet→GFP glows
 
 
I13453+RBS+[http://partsregistry.org/Part:BBa_C0072 C0072]+TT+K091105+E0240
 
 
 
Test Tet and Mnt repression:                     
 
                                       
 
Some promoter→TetR+Mnt+t+t+pMnt/Tet→GFP does not glows
 
 
I13453+RBS+C0040+C0072+TT+K091105+E0240
 
 
add aTc does not glow
 

Latest revision as of 15:27, 18 July 2008

This are our ideas of how to test the four hybrid promoters for the XOR gate.


Test pLuxR/cI


Test pMnt/Lac


Test pLsrA/cI


Test pLasR/cI


Test Mnt/Tet promoter


Overlapping Parts And intermediates


LuxR/cI Lac/Mnt LsrR/cI LasR/cI Mnt/Tet
RBS+cI(+TT) RBS+cI(+TT)
RBS+Mnt(+TT) RBS+Mnt(+TT)