Part:BBa_K613010:Experience

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Applications of BBa_K613010

Experience of the ITB_Indonesia iGEM 2015 team.

The iGEM 2015 ITB Indonesia used this promoter and RBS to express BBa_K653000 for rhamnolipid production in E. coli BL21(DE3). After IPTG induction, the separated supernatant was tested for surfactant activity.
Supernatant from empty cell, either induced or not, had no / little surfactant activity. The uninduced transformant had little activity, but the induced transformant had higher surfactant activity.
This shows that the promoter can be induced for BBa_K653000 expression.

Characterization of Designed Variants by EPFL 2011

For each family, we tested the randomers and the designed variants separately. To characterize the promoter strengths, we used RFP as the reporter gene and used a platereader to test for fluorescence during and after induction with IPTG.

The six designed T7 promoter variants are named as a function of their predicted promoter efficiency, relative to the wildtype. For example, T7 14 has a predicted efficiency of 14% compared to the consensus T7 promoter, whereas T7 111 is predicted to be 111% more efficient than the wildtype. In the chart above, each of the designed promoter variants for both the T7 with and without the lac operator are arranged in increasing predicted efficiency. Contrary to our expectation, some variants that were designed to have a lesser efficiency than the wildtype (e.g. T7 54) seem to have a much higher strength (as measured by fluorescence at saturation, normalized by the optical density). The data for this graph was produced in triplicate, so the error bar represents the standard error across those three measurements.

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