Difference between revisions of "Part:BBa K526000"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K526000 short</partinfo>
 
<partinfo>BBa_K526000 short</partinfo>
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'''fim inversion system'''
 
'''fim inversion system'''
  
 
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.  
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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).
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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).
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[[Image:SKL7.jpg|300px]]
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[[Image:SKL7.jpg|250px]]       
  
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'''Light Receptor'''
 
  
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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.
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'''Light Receptor'''
  
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[[Image:SKL8.jpg|300px]]
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Pompc is a promoter sequence that is regulated by the hybrid light receptor, Cph8. Unfortunately we need to have the continous presence of darkness in order to induce the gene expression. Maintenance of complete darkness (especially on an industrial scale) for a longer duration is impossible. Hence, many researchers tried to introduce an invertor (usually the lambda repressor, cI). But even this system demands the continous presence of light which is again practically difficult. Hence, we propose a light mediated pulse inducer using the fimE processor. <br/><br/>
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'''A Novel Light Based Pulse Induction System'''
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<p>We made an attempt to introduce a light based pulse induction system using the 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. The system would be permanently on when all of the promoter is inverted indicating need for no further inducer. This system would also ensure more tight control of target gene expression. This combination would be potentially useful for toxic gene expression as the basal level of the toxic gene would be kept as low as possible. </p>
  
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'''Solution'''
 
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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.
 
  
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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.
 
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[[Image:SKL6.jpg|300px]]
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 04:27, 6 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 hybrid light receptor, Cph8. Unfortunately we need to have the continous presence of darkness in order to induce the gene expression. Maintenance of complete darkness (especially on an industrial scale) for a longer duration is impossible. Hence, many researchers tried to introduce an invertor (usually the lambda repressor, cI). But even this system demands the continous presence of light which is again practically difficult. Hence, we propose a light mediated pulse inducer using the fimE processor.

A Novel Light Based Pulse Induction System

We made an attempt to introduce a light based pulse induction system using the 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. The system would be permanently on when all of the promoter is inverted indicating need for no further inducer. This system would also ensure more tight control of target gene expression. This combination would be potentially useful for toxic gene expression as the basal level of the toxic gene would be kept as low as possible.


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]