Difference between revisions of "Part:BBa K526000"
Line 4: Line 4:
  
 
fimE is a invertase,by function,that is capable of inverting any DNA sequence that is flanked by fimE recognition site.  
 
fimE is a invertase,by function,that is capable of inverting any DNA sequence that is flanked by fimE recognition site.  
−
When fimE recognition sequences are flanked to a promoter element we can ensure that there is no leaky expression as the promoter can be maintained in the opposite orientation till the presence of inducer. Thus, fim inversion promoters ensures that there is no leaky expression (Fig 1).
+
When fimE recognition sequences are flanked to a promoter element we can ensure that there is no leaky expression as the promoter can be maintained in the opposite orientation till the presence of inducer. Thus, fim inversion promoters ensures that there is a tight regulation of target gene expression (Fig 1).
−
 
+
−
[[Image:SKL7.jpg|250px]]          [[Image:SKL8.jpg|300px]]
+
  
 +
[[Image:SKL7.jpg|250px]]       
  
  
 
'''Light Receptor'''
 
'''Light Receptor'''
  
−
Pompc is a promoter sequence that is regulated by the famous hybrid light receptor Cph8. Cph8 mediated regulation of gene expression suffers from a high leaky expression (Fig 2).
+
Pompc is a promoter sequence that is regulated by the famous hybrid light receptor Cph8.  
−
 
+
  
 
'''Solution'''
 
'''Solution'''
Line 19: Line 17:
 
fim inversion promoter system is used to provide a tight control of gene expression. On the other hand, light inducible gene expression system suffers from a high leaky expression (Fig 3). Hence, we made an attempt to reduce the leaky level of the light mediated gene expression systems by using a combination of light receptor and fimE processor. In this biobrick part, we have made an attempt to regulate the expression of fimE based on light so that we could precisely turn on a previously uninduced gene . This system would ensure more tight control of target gene expression. This combination would be potentially useful for toxic gene expression.  
 
fim inversion promoter system is used to provide a tight control of gene expression. On the other hand, light inducible gene expression system suffers from a high leaky expression (Fig 3). Hence, we made an attempt to reduce the leaky level of the light mediated gene expression systems by using a combination of light receptor and fimE processor. In this biobrick part, we have made an attempt to regulate the expression of fimE based on light so that we could precisely turn on a previously uninduced gene . This system would ensure more tight control of target gene expression. This combination would be potentially useful for toxic gene expression.  
  
−
Contrary to the expectation, a combination of light inducible system and fim inversion system lead to a constitutive gene expression (Fig 3). Hence, we hyphothesize that optimizing the RBS would help avoid this basal expression and favor a tight regulation of gene expression.
+
Contrary to the expectation, a combination of light inducible system and fim inversion system lead to a constitutive gene expression (Fig 2). Hence, we hyphothesize that optimizing the RBS would help avoid this basal expression and favor a tight regulation of gene expression.
 
[[Image:SKL6.jpg|300px]]
 
[[Image:SKL6.jpg|300px]]
  

Revision as of 06:45, 3 October 2011

Pompc promoter coding fimE fim inversion system

fimE is a invertase,by function,that is capable of inverting any DNA sequence that is flanked by fimE recognition site. When fimE recognition sequences are flanked to a promoter element we can ensure that there is no leaky expression as the promoter can be maintained in the opposite orientation till the presence of inducer. Thus, fim inversion promoters ensures that there is a tight regulation of target gene expression (Fig 1).

SKL7.jpg


Light Receptor

Pompc is a promoter sequence that is regulated by the famous hybrid light receptor Cph8.

Solution

fim inversion promoter system is used to provide a tight control of gene expression. On the other hand, light inducible gene expression system suffers from a high leaky expression (Fig 3). Hence, we made an attempt to reduce the leaky level of the light mediated gene expression systems by using a combination of light receptor and fimE processor. In this biobrick part, we have made an attempt to regulate the expression of fimE based on light so that we could precisely turn on a previously uninduced gene . This system would ensure more tight control of target gene expression. This combination would be potentially useful for toxic gene expression.

Contrary to the expectation, a combination of light inducible system and fim inversion system lead to a constitutive gene expression (Fig 2). Hence, we hyphothesize that optimizing the RBS would help avoid this basal expression and favor a tight regulation of gene expression. SKL6.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 750
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 750
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 750
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 750
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 750
  • 1000
    COMPATIBLE WITH RFC[1000]