Team:Amsterdam/Labwork/Characterization

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Under construction
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=Characterization=
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So in the end, we got two working assembly’s.
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==Cpn10==
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The first is a construct with lacI, strong RBS and Cpn10. In our E.coli lacI is always present so the promoter is always on. Cpn10 was selected, because it is a chaperone that’s works together with Cpn60 and helps the folding of proteins at lower temperatures.
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We don’t suspect any influence of Cpn10 on the growth curves, because it is the smaller one of the two chaperones. Unfortunately we didn’t get CPN60 in a construct.
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==CspC==
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The second construct is lacI, strong RBS and CspC. CspC is known for its influence on survivability whit a Freeze/thaw cycle. This means that you freeze the bacteria, than thaw them, freeze them, thaw them, etc and look at the survival rate. So we don’t expect effect on the growth rate, at lower temperatures. But we do expect a higher survival rate when you freeze and thaw it.
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==Growth curves==
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[under construction]
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In figure 1 u see the specific growth rate against the temperature. We expected that if our genes would have any effect, it would be a higher specific growth rate at low temperatures (4 till 30°C), and no effect on the specific growth rate at 37°C (the optimal temperature for E.coli). As you can see in the graph this isn’t visible. So our Cpn10, and CspC have no significant effect on the growth rate.
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{{:Team:Amsterdam/Footer}}
{{:Team:Amsterdam/Footer}}

Revision as of 14:07, 20 September 2011

Contents

Characterization

So in the end, we got two working assembly’s.

Cpn10

The first is a construct with lacI, strong RBS and Cpn10. In our E.coli lacI is always present so the promoter is always on. Cpn10 was selected, because it is a chaperone that’s works together with Cpn60 and helps the folding of proteins at lower temperatures. We don’t suspect any influence of Cpn10 on the growth curves, because it is the smaller one of the two chaperones. Unfortunately we didn’t get CPN60 in a construct.

CspC

The second construct is lacI, strong RBS and CspC. CspC is known for its influence on survivability whit a Freeze/thaw cycle. This means that you freeze the bacteria, than thaw them, freeze them, thaw them, etc and look at the survival rate. So we don’t expect effect on the growth rate, at lower temperatures. But we do expect a higher survival rate when you freeze and thaw it.

Growth curves

[under construction]

In figure 1 u see the specific growth rate against the temperature. We expected that if our genes would have any effect, it would be a higher specific growth rate at low temperatures (4 till 30°C), and no effect on the specific growth rate at 37°C (the optimal temperature for E.coli). As you can see in the graph this isn’t visible. So our Cpn10, and CspC have no significant effect on the growth rate.