Part:BBa_K2078002:Experience
Contents
Applications of BBa_K2078002
Experience of CLSB-UK-2016 iGEM team with Part:BBa_K2078002
Experimental Design
This consensus promoter was assembled with the reporter protein, blue-purple chromoprotein AmilCP. The construct was carried on the pSB1C3 plasmid for expression and amplification in E-Coli. The plasmid also contained chloramphenicol acetyltransferase for resistance during plating (CmR).
Observations and Inferences
Our E.coli grew slower than other transformants and we were also unable to recover as much plasmid by miniprep. The use of this strong consensus promoter slows down expression times of the AmilCP Part:BBa_K592025 in E-Coli. This image shows a pellet from 5ml LB broth with transformed E.coli and confirms that the part is expressed, but the amount of pellet was much less than we would normally expect to get from 5ml.
The delay in E-Coli growth is also shown by this 24hour streak plate. The chromoprotein is only just becoming visible suggesting that such a strong consensus sequence slows down the expression time of the part. E-Coli also grows less quickly when transformed with the part. This is thought to be due to the high binding strength of RNAPol to the consensus promoter sequence which slows down the rate of transcrption
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Synechocystis 6803 was successfully transformed with the part but no colour could be seen.
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