Difference between revisions of "Part:BBa K784001:Experience"
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We have used the part inside a BL21 strain of E.coli which has an endogenouse PET system. | We have used the part inside a BL21 strain of E.coli which has an endogenouse PET system. | ||
In the presence of IPTG inducer, T7 RNAP is expressed and activates the part promoter which expresses the xyIE gene. <br> | In the presence of IPTG inducer, T7 RNAP is expressed and activates the part promoter which expresses the xyIE gene. <br> | ||
Revision as of 15:40, 23 September 2012
Experience of BBa_K784001
We have used the part inside a BL21 strain of E.coli which has an endogenouse PET system.
In the presence of IPTG inducer, T7 RNAP is expressed and activates the part promoter which expresses the xyIE gene.
The graph below presents the results of the experiment described. The absorbance at 380nm for each of the different concentrations of the inducer in a range of 0.5 μM to 500 μM on a logarithmic scale. The absorbance was calculated via different dilutions of the samples: dilutions by 0.5, by 0.25, and by 0.1. The error bars represent the processing of the data collected. The line at the bottom of the graph represents the basal level according to the control result- no IPTG induction.
As can be seen, the graph shows a small tendency to rise, but it's not clear enough. There are several explanations, one is that saturation has been achieved from lowest concentration measured, which shows both that the assay is highly sensitive and that a small amount of polymerase is enough to cause translation. The experiment should be repeated with either lower concentrations of IPTG or Catechol. The line at the bottom of the graph represents the basal level according to the control result- no IPTG induction.
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