Difference between revisions of "Part:BBa K526000"

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).

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]

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 '''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.
+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/>
 '''A Novel Light Based Pulse Induction System'''
+  <p>We made an attempt to introduce a light based pulse induction system using the fimE processor.
- 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.
+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>
-Contrary to the expectation, a combination of light inducible system and fim inversion system led to a constitutive gene expression. Hence, we hyphothesize that optimizing the RBS of fimE processor would help avoid this basal expression and favor a tight regulation of gene expression.