Team:Imperial College London/M3/Genetic1

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When sufficient protein is being produced and encapsulation is complete, temperatures will be increased to 42°C. This results in the de-repression of the pLambda promoter and restriction enzymes DpnII and TaqI will be produced. Dam methylase will not be able to protect the E. coli DNA from being cut and genome deletion would occur. This causes cell death and destruction of possible pathogenicity from E. coli.
When sufficient protein is being produced and encapsulation is complete, temperatures will be increased to 42°C. This results in the de-repression of the pLambda promoter and restriction enzymes DpnII and TaqI will be produced. Dam methylase will not be able to protect the E. coli DNA from being cut and genome deletion would occur. This causes cell death and destruction of possible pathogenicity from E. coli.
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==Principle==
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{| class="wikitable" border="1"
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|- style="background:blue; color:white"
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!  Temperature
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!  DpnII, TaqI (Restriction enzymes)
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!  Dam (Methylation)
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|- style="background:blue; color:white"
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|  Below Threshold
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|  OFF
 +
|  ON (Protects)
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|- style="background:blue; color:white"
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|  Above Threshold
 +
|  ON
 +
|  ON (Not enough to protect DNA)
 +
|}
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 +
{| style="color:#CCC; background-color:#3D5089;" cellpadding="6" cellspacing="0" border="1"
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! Temperature
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! DpnII, TaqI (Restriction enzymes)
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! Dam (Methylation)
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|- style="color:#333; background-color:#BCD2EE;" cellpadding="6" cellspacing="0" border="1"
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<!----------------------------------------biobricks---------------------------------------->
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|Below Threshold
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|  OFF
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|  ON (Protects)
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|- style="color:#333; background-color:#BCD2EE;" cellpadding="6" cellspacing="0" border="1"
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|  Above Threshold
 +
|  ON
 +
|  ON (Not enough to protect DNA)
 +
|}
 +
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Latest revision as of 14:22, 11 September 2009

Contents

M3 Genetic Circuit

Video

Genetic Circuit

M3.fgc1.PNG

Elaboration

This is the genetic circuit for thermally induced genome deletion (Module 3).

This construct has two components. The first component controls the production of restriction enzymes DpnII and TaqI. The second component will ensure the production of Dam methylase, which protects the DNA.

Protein production and encapsulation occurs 28°C. In addition, protein cI is produced constitutively at 28°C. It represses the lambda cI promoter, hence only basal amounts restriction enzymes will be produced. At the same time, the production of Dam methylase will protect the E. coli DNA by methylation, and hence prevent cell death. Genome deletion would not occur.

When sufficient protein is being produced and encapsulation is complete, temperatures will be increased to 42°C. This results in the de-repression of the pLambda promoter and restriction enzymes DpnII and TaqI will be produced. Dam methylase will not be able to protect the E. coli DNA from being cut and genome deletion would occur. This causes cell death and destruction of possible pathogenicity from E. coli.

Principle

Temperature DpnII, TaqI (Restriction enzymes) Dam (Methylation)
Below Threshold OFF ON (Protects)
Above Threshold ON ON (Not enough to protect DNA)
Temperature DpnII, TaqI (Restriction enzymes) Dam (Methylation)
Below Threshold OFF ON (Protects)
Above Threshold ON ON (Not enough to protect DNA)


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